Vous êtes sur la page 1sur 590

CAT No.

8040883

Operator's Manual

VITRO

' Ortho:Clinical Diagnostics


a ^o&wton^flo^HiOH company

Export authorized under general license GTDA (General Technical Data Available) IMPORTANT The information contained herein is based on the experience and knowledge relating to the subject matter gained by Ortho-Clinical Diagnostics, Inc. prior to publication. No patent license is granted by the information. Ortho-Clinical Diagnostics, Inc. reserves the right to change this information without notice, and makes no warranty, express or implied, with respect to the information.. The company shall not be liable for any loss or damage including consequential or special damages, resulting from the use of this information, even if loss or damage is caused by its negligence or other fault.

VITROS is a trademark of Ortho-Clinical Diagnostics.

2004 Ortho-Clinical Diagnostics, Inc. All rights reserved.

2004-03-30

About the VITROS DT II System Binder


This binder contains all you need to know to analyze samples on the VITROS DT60 II Chemisty System and the accompanying VITROS DTSC II and VITROS DTE II Modules.

Operator's Manual
This section provides general information about how to operate and maintain the VITROS DT II System. Topics covered are: Operating Instructions Coronary Risk Classification (CRC) and Derived Tests Calibration Instrument Care and Cleaning Quality Control Options Troubleshooting Instrument Status Messages Coded Warning Messages Installation and Site Specifications Warranty In writing the Operator's Manual we strove to achieve multiple goals in meeting your needs. The Operator's Manual has some sections (1, 6, and 7-12) that are primarily informative. These are formatted vertically, making it easier to read these sitting down or at a desk. Sections 2-5 and 1 3 are considered functional, for daily use while you are operating the analyzer, and are oriented horizontally.

Rev. 2004-03-30

VITROS DT II System Binder VITROS DT II System

I
|

VITROS Chemistry Products DT Instructions for Use Manual


This separate manual provides information on the specific chemistries that the DT II System analyzes. It includes the following information for each chemistry: Intended Use Summary and Explanation of the Test Principles of the Procedure Test Type and Conditions Warnings and Precautions Reagents Specimen Requirements Testing Procedure Calibration Quality Control Expected Values and Reporting Units Limitations of the Procedure Performance Characteristics

iv

VITROS DT II System Binder VITROS DT l| System

Rev. 2004-03-30

Revision History

Revision Date
2004-03-30

Description
About the VITROS DT II System Binder iii-iv, updated to reflect moving Instructions for Use to separate manual Revision History v-viii, updated to reflect current documentation List of Revised Pages ix-x, updated to reflect current documentation Table of Contents xi-xiv, updated to reflect current documentation Chapter 1, page 1 - 1 , removed reference to Hb Chapter 2, page 2-19, 2-23 and 2-25, inserted text for tracking error Chapter 4, page 4-3, updated "When to Calibrate" section Chapter 4, page 4-3, removed reference to Hb Chapter 4, page 4-11, removed section "Preparing DT Hb Calibrators" Chapter 4, page 4-13 and 4-15, inserted text for tracking error Chapter 6, page 6-3, removed section "Preparing DT Hb controls" Chapter 8, page 8-3, added "Unexpected Results" to section title, added bullet for calibration failures Chapter 8, page 8-4, In 'Possible Causes" extracted text from third bullet to emphasize importance, added bullet for overfilled slide disposal box Chapter 8, page 8-4, In "if You Suspect a Tracking Error" added text to review and confirm previous results Chapter 8, page 8-4, In "Important Points to Remember" underlined text for emphasis Chapter 9, page 9-8, added Important note to prevent tracking error Chapter 1 0, page 10-1 and 10-3, added tracking error text Chapter 1 0, page 10-2 and 10-20, removed reference to Hb Chapter 10, page 10-10, added tracking error text

Rev. 2004-03-30

Operator's Manual VITROS DT II System

Revision Date
2003-10-01

Description
Note: Formatting has changed but only pages with content changes are labeled 2003-10-01. Added "For in vitro Diagnostic Use" Replaced "Methodology Sheets" with "Instructions for Use" About, pages iii-iv, edited to reflect current documentation Revision History, v-vii, added update information List of Revised Pages, viii, updated to include all pages Table of Contents, xi-xii, updated Introduction, page 1-1 Introduction, pages 1-7-1-8 Introduction, pages 1 -7 and 1 -9; removed DT from VITROS Micro Tips Section 2.2.1, page 2-5 Section 2.2.1, page 2-7, updated the power socket illustration Section 2.2.1, page 2-27, changed J&JCD to OCD Se tion 6.10, page 6-13 Section 7.2, page 7-3, changed J&JCD to OCD Section 7.2, page 7-7, changed J&JCD to OCD Section 8.2, page 8-3-8-6, replaced information with a reference; changed paging to 8-3-8-4 Section 9.1, page 9-5 Section 10.1.8, page 10-19 Revised Instructions for Use: ALB DT ALT DT AST DT BUN DT CHE DT CKDT Cl- DT CREA DT FeDT LAC DT LDH DT Mg DT Na + DT NBIL DT N H , DT THEO DT TBIL DT TPDT UrCr DT

VI

Operator's Manual VITROS DT II System

Rev. 2004-03-30

Revision Date
2003-08-11

Description
Revised Instructions for Use for: AMYL DT CO 2 DT GGT DT GLU DT HDLC DT (Using the VITROS DT HDL Cholesterol Kit) HDLC DT (Using the VITROS DT Micro HDL Cholesterol Kit) Li DT URIC DT Removed: Anion Gap Calculation Supplement, C-363 Coronary Risk Classification Supplement, C-359 Globulin and Albumin_Globulin Calculations Supplement, C-360 VLDL, LDL, and CHOLJHDLC Ratio Calculations Supplement, C-362 New Format. New organization and sections consistent with IVD Directive. Revised Instructions for Use: ALKP DT Ca DT CHOL DT CKMB DT CRSC DT K+ DT LI PA DT PHOS DT TRIG DT Update the Methodology Section to include revision to Test Methodology: CK2003FEB01 Revised Table of Contents to include Test Methodology Sheets. Revised Section 5, pages 1-46, deleted all references to cleaning and charging the VITROS DT Pipette. Refer to the VITROS DT Pipette User's Guide for information on cleaning and charging the DT Pipette. Section 5, pages 1-10, minor text and format changes. Section 5.1, page 3, deleted reference to the daily cleaning of the DT Pipette. Section 5.3, pages 19-26, removed. Section 5.4, pages 27-34, removed. Updated Methodology Section to include Test Methodologies: NBIL 2001 SEP24, TBIL 2001 SEP24. Updated Methodology Section to include Test Methodologies: GGT 8/99, CK 2000SEP21, CREA 2000SEP21. Reflects company name change to Ortho-Clinical Johnson Company. Diagnostics, a Johnson &

2003-04-30

2OO3FEB 10/01

11/00 1/99 4/96

Reprinted for the introduction of J&JCD VITROS Chemistry Systems trademark nomenclature.

Rev. 2004-03-30

Operator's Manual VITROS DT II System

VII

Revision Date
5/95

Description
Reprint of all pages. Trade dress changes. Style changes for consistency. Section 2.2.1, pages 7-1 7, changes made to reflect new pipette information. Section 2.2.2, page 23, item 6, new information added. Section 4.2, minor text changes. Section 4.4, page 13, pipette graphic updated. Section 4.4, page 15, step 4, minor text changes. Section 5.3, pages 19-25, new information on cleaning the DT Pipette. Section 5.4, pages 27-33, heading changed to Charging the DT Pipette and new information added. Section 5.5, page 35, clean cloth changed to cotton swab. Section 8.2, page 3, new pipette troubleshooting information added. Section 8.3, Analyzer Tracking Errors section added. Section 11.1, page 2, item 3, reference to Electrical Requirements added. Section 11.2.3, page 5, rewrite. Section 11.3, page 6, changes in headings and rewrite of item 2 under 11.3.1 and 11.3.2. Reprint of all pages. Table of Contents undated to reflect derived tests in Section 3 and switch of subheads in Section 11. Section 2, page 13, second bullet under Ektachem DTE pipette removed. Section 3, style changes to comply with other subheads. Section 3, pages 11 and 13, md/dl changed to mg/dl. Section 3, pages 18-36, new pages added as per derived test development. Section 4.3.2, page 11, second bullet changed. Section 4, page 1 7, IMPORTANT NOTE changed. Section 6.4.2, page 3, second bullet changed. Section 7, pages 4-10, new pages added as per derived test development. Section 11.3.1 and 11.3.2, heading corrections. Updated Sections 3 & 7 to include new features. Revised tab for Section 3 to include both CRC and Derived Tests. Revised Section 3 in Table of Contents to reflect additions. First release.

9/93 2/92

1/92

9/91

Operator's Manual VITROS DT II System

Rev. 2004-03-30

List of Revised Pages


Each page in your manual should be at the date listed below: Chapter or Section
Title Page/Copyright About Revision History List of Revised Pages Table of Contents (book) Introduction iii-iv v-viii ix-x xi-xiv 1-1 1-2-1-6 1-7-1-9 1-10 2-1-2-4 2-5 2-6 2-7 2-8-2-18 2-19 2-20-2-22 2-23 2-24 2-25 2-26 2-27 2-28-2-34 3-1-3-36 4-1-4-2 4-3 4-4-4-10 4-11 4-12 4-13 4-14 4-15 4-16-4-22 5-1-5-30 6-1-6-2 6-3 6-4-6-12 6-13 6-14 7-1-7-2 7-3 7-4-7-6 1-7 7-8-7-10 8-1-8-2 8-3-8-4

Page Number(s)/Portrait (P) or Landscape (L)


P P P P P P P P P L L L L L L LL L L L L L L L L L L L L L L L L L P P P

Revision Date
Rev. 2004-03-30 Rev. 2004-03-30 Rev. 2004-03-30 Rev. 2004-03-30 Rev. 2004-03-30 Rev. 2004-03-30 5/95. Reprinted 1/99 Rev. 2003-10-01 5/95. Reprinted 1/99 5/95. Reprinted 1/99 Rev. 2003-10-01 5/95. Reprinted 1/99 Rev. 2003-10-01 5/95. Reprinted 1/99 Rev. 2004-03-30 5/95. Reprinted 1/99 Rev. 2004-03-30 5/95. Reprinted 1/99 Rev. 2004-03-30 5/95. Reprinted 1/99 Rev. 2003-10-01 5/95. Reprinted 1/99 5/95. Reprinted 1/99 5/95. Reprinted 1/99 Rev. 2004-03-30 5/95. Reprinted 1/99 Rev. 2004-03-30 5/95. Reprinted 1/99 Rev. 2004-03-30 5/95. Reprinted 1/99 Rev. 2004-03-30 5/95. Reprinted 1/99 Rev. 10/01 5/95. Reprinted 1/99 Rev. 2004-03-30 5/95. Reprinted 1/99 Rev. 2003-10-01 5/95. Reprinted 1/99 5/95. Reprinted 1/99 Rev. 2003-10-01 5/95. Reprinted 1/99 Rev. 2003-10-01 5/95. Reprinted 1/99 5/95. Reprinted 1/99 Rev. 2004-03-30

Operating Instructions

CRC Calibration

Instrument Care & Cleaning Quality Control

Options

P P P P P P P

Troubleshooting

Rev. 2004-03-30

Operator's Manual VITROS DT II System

IX

Chapter or Section Status Messages

Page Number(s)/Portrait (P) or Landscape (L) 9-1-9-4 9-5 9-6-9-7 9-8 10-1-10-3 10-4-10-9 10-10 10-11-10-18 10-19 10-20 10-21-10-22 11-1-11-6 12-1-12-2 13-1-13-3 13-4-13-8 P P P P P P P P P P P P P L P

Revision Date 5/95. Reprinted 1/99 Rev. 2003-10-01 5/95. Reprinted 1/99 Rev. 2004-03-30 Rev. 2004-03-30 5/95. Reprinted 1/99 Rev. 2004-03-30 5/95. Reprinted 1/99 Rev. 2003-10-01 Rev. 2004-03-30 5/95. Reprinted 1/99 5/95. Reprinted 1/99 5/95. Reprinted 1/99 5/95. Reprinted 1/99. 5/95. Reprinted 1/99

Coded Warning Messages

Installation and Site Spec Warranty Log Sheets

Operator's Manual VITROS DT II System

Rev. 2004-03-30

Table of Contents
About the VITROS DT II System Binder Operator's Manual VITROS Chemistry Products DT Instructions for Use Manual Revision History List of Revised Pages Table of Contents 1.1 1.2 General Description Equipment Features 1.2.1 Keyboard and Functions 1.2.2 Display Panel 1.2.3 Printer and Results Printout Principles of Operation: VITROS DT60 II Chemistry System 1.3.1 Slide Identification 1.3.2 Slide Spotting 1.3.3 Incubation and Readout Principles of Operation: VITROS DTE II Module 1.4.1 Slide Identification 1.4.2 Slide Spotting 1.4.3 Incubation and Readout Principles of Operation VITROS DTSC II Module 1.5.1 Slide Identification 1.5.2 Slide Spotting 1.5.3 Incubation and Readout Supplies and Supply Handling and Storage 1.6.1 Storage Temperature Requirements 1.6.2 VITROS DT Slides 1.6.3 VITROS DT and DTE Pipettes and Micro Tips 1.6.4 VITROS DT Calibrator Kits 1.6.5 ViTROS DT Control I and DT Control II 1.6.6 VITROS DT Reference Fluid 1.6.7 VITROS DTE Dual-Sample Cups.. 1.6.8 Printer Paper 1.6.9 VITROS DT Accessory Kit* iii iii iv v ix xi 1-1 1-2 1-2 1-3 1-3 1-3 1-3 1-3 1-4 1-5 1-5 1-5 1-5 1-6 1-6 1-6 1-6 1-7 1-7 1-7 1-7 1-7 1-8 1-8 1-8 1-8 1-9

Section 1 Getting To Know Your VITROS DT II System

1.3

1.4

1.5

1.6

Section 2 Operating Instructions

2.1 2.2

2.3

Start-up Procedure Testing Procedure 2.2.1 Pipetting Techniques 2.2.2 Steps for Analysis on the VITROS DT II System .... Calibration Data Module and Chemistry Language Module 2.3.1 Description of Calibration Data Module

2-1 2-5 2-5 2-19 2-27 2-27

Rev. 2004-03-30

Operator's Manual VITROS DT II System

xi

2.3.2 2.3.3 2.4 2.5

Description of Calibration Language Module 2-27 How to Install a New Calibration Data Module (CDM) and a Chemistry Language Module (CLM) .2-29 Normal Shutdown Procedure 2-33 Emergency Shutdown 2-33 3-1 3-3 3-13 3-21 3-21 3-23 3-27 3-33 4-1 4-3 4-5 4-7 4-11 4-13 5-3 5-5 5-19 5-21 5-21 5-23

Section 3 Coronary Risk Classification (CRC)

3.1 CRC Overview 3.2 Entering Data for Coronary Risk Classification 3.3 Printing CRC Results Derived Tests 3.4 Overview 3.5 GLOB and A/G 3.6 Lipid Calculation (VLDL, LDL and CHOL/HDLC Ratio) 3.7 Anion Gap (AGP) 4.1 4.2 4.3 Why You Need to Calibrate When to Calibrate How to Calibrate 4.3.1 Preparing VITROS DT Calibrators 4.3.2 Entering the Calibration Mode Calibration Daily Cleaning Weekly Cleaning Other Cleaning: VITROS DT60 II Chemistry System FORSHead '... .. Paper Loading ,;;: 5.4.1 Removing the Paper 5.4.2 Inserting Paper '.

Section 4 Calibration

4.4

Section 5 Instrument Care and Cleaning

5.1 5.2 5.3 5.4

Section 6 Quality Control

6.1 6.2 6.3 6.4 6.5 6.6

6.7

6.8

What Is Quality Control? What Are Controls? How Often Should Controls Be Run? How to perform a quality control test 6.4.1 Preparing Lyophilized Controls Analyzing the Controls How to Record the Results 6.6.1 Recording Quality Control Results on a Log 6.6.2 Recording Quality Control Results on a Graph Interpreting Control Results 6.7.1 Interpreting the Control Results from the Log 6.7.2 Interpreting the Control Results from the Graph Establishing Your Own Control Ranges 6.8.1 How to Calculate the Mean 6.8.2 Variables to Consider in Establishing the Mean

6-1 6-1 6-1 6-2 6-2 6-3 6-4 6-4 6-5 6-6 6-6 6-6 6-7 6-8 6-9

XII

Operator's Manual VITROS DT II System

Rev. 2004-03-30

6.8.3 Calculating the Standard Deviation 6.8.4 How to Calculate a Standard Deviation 6.9 Quality Control Troubleshooting Chart 6.10 Factors to Consider When Your Results Are Out of Range

6-9 6-10 6-12 6-13

Section 7 Options

7.1 7.2

7.3 7.4

How to Run Options Options for the VITROS DT60 II Chemistry System 7.2.1 Serial Communication Options for the DT60 II System Options for the VITROS DTE II Module Options for the VITROS DTSC II Module

7-1 7-1 7-8 7-9 7-10

Section 8 Troubleshooting

8.1 8.2 8.3

General Troubleshooting Pipette Troubleshooting Unexpected Results (Analyzer Tracking Errors)

8-1 8-3 8-3

Section 9 Instrument Status Messages Section 10 Coded Warning Messages

9.1

Status Messages

9-1

10.1 Coded Warning Messages 10.1.1 Calibration (C) 10.1.2 Data Storage (D) 10.1.3 Electrometer (E) 10.1.4 Instrument Function (F) 10.1.5 Temperature (H) 10.1.6 Communications (N) 10.1.7 Reflectometer (R) DT60 II System 10.1.8 Reflectometer (L) DTSC II Module 11.1 Installation 11.2 Site Specifications 11.2.1 Space Requirements 11.2.2 Environmental Requirements (Temperature, Humidity, and Altitude) 11.2.3 Electrical Requirements 11.2.4 Refrigerator and Freezer Space 11.3 Moving the Analyzer 11.3.1 Relocation Outside the Office 11.3.2 Relocation Within the Office 12.1 New Equipment Warranty VITROS DT II System 12.2 New Accessory Warranty VITROS DT Pipette and VITROS DTE Pipette

10-1 10-1 10-4 10-6 10-10 10-14 10-16 10-18 10-19 11-1 11-3 11-3 11-4 11-5 11-6 11-6 11-6 11-6 12-1 12-2

Section 11 Installation and Site Specifications

Section 12 Warranty

Rev. 2004-03-30

Operator's Manual VITROS DT II System

XIII

Section 13 Log Sheets

CALIBRATION LOG 13-1 SERVICE LOG 13-2 TEST/REAGENT LOG 13-3 VITROS DT II System Maintenance Log 13-4 VITROS DT II System Quality Control Log 13-5 Levey-Jennings Quality Control Chart for VITROS DT II System.13-6

Section 14 Instructions For Use

xiv

Operator's Manual VITROS DT II System

Rev. 2004-03-30

Getting To Know Your VITROS DT II System


This manual is designed to familiarize you with the operation of your VITROS DT60 II Chemistry System and, if applicable, your ViTROS DTE II and VITROS DTSC II Modules. We suggest that the operator become familiar with the information in this manual prior to operating the analyzer and the associated modules.

1.1 General Description


Intended Use

For in vitro diagnostic use. The VITROS DT60 II System uses VITROS DT Slides to perform a number of discrete clinical tests on serum or plasma specimens. All reactions needed for a single quantitative measurement take place within the multilayered analytical element of the slide. A slide is used once for a single patient test and is then discarded. The unique properties of these slides eliminate the need to store, mix, and dispose of liquid reagent chemicals and permit reliable analyses with a very small patient sample. All processes performed by the analyzer are controlled by a selfcontained microcomputer. You communicate with the microcomputer through the keyboard; it communicates with you through messages that appear on the display screen and printouts.

TABLE 1 Summary of System Characteristics


.and

^m^iM^MstC:

h. ::! I l l : :Jhr:

: the specific Instructions;for Use^ta;rrrore: infofrrfaiti(Jinr; 'Specimen Sample: - : : : m : "" :v ;: ):'!':: ""% N#i::::;:;; : :.n ; C i i H J Serum or|p)asma* ; : ';S.f-wi:;H-|Vi; i -^XM^/ Sample Size:
:;

-^r^
:

: y ' -^ ^'un^&S^,
L - v i i J JJiinii..

:1 ^'^
M
;

; 1 ; Q ( J L p e r t e s t r T " ; ^ : : ; : ; / : :: : :

per -lest JResutfc

VITR(3iS:PT60 II Chemistry VlTRQfilpE II Module: A dJfc iModule:


^ f K W | > M g h p u t Rate:" " '^^iW^Bttlu; I ^"'1 DTOpli : l-i|eni^try:Systern: i;: ApproximiSlf.feS tests per hour 0 I E i r M o | ( u ] e : i'Approxiniately 15 tests^erihour -M : y' : : ' ^ ^ M l ^ = ^^fflGscJWlijt^tiK" -t 5 f^M-1^!^1^ -hour:|-:i:':.:. : .

* Appropriate anticoagulants for plasma specimens are suggested in the test Instructions for Use

Rev. 2004-03-30

Operator's Manual VITROS DT II System

1-1

1.2 Equipment Features 1.2.1 Keyboard and Functions


The keyboard is used to issue commands to the analyzer's microcomputer and to enter necessary data. Instructions for using the keyboard are provided in this section. To protect the analyzer against damage from spills and to make the equipment easier to clean, the keyboard consists of a pressuresensitive pad rather than actual keys. An audible tone sounds shortly after a key is pressed. If no tone sounds, you have to apply more pressure to the center of the key. A double tone sounds if you attempt to enter information that is incorrect.

patient ID in progress complete

print service mode cal mode delete test chemistry select enter

' clear .shift

SHIFT:

NUMERIC KEYS:

Used to perform all functions shown in red on your keyboardtest in progress, service mode, and delete test. Use the shift key first, then proceed with your second key selection.
CLEAR:

Used in entering numeric data, such as patient identification numbers, and in identifying calibrators used in the calibration procedure.
ENTER:

Erases data from the display panel. TEST IN PROGRESS/COMPLETE: Test in Progress: (Press SHIFT key.) Used to display tests that are currently in the incubator of the DT60 II System and any active tests in the DTE II or DTSC II Module. Test Complete: Used to display the results of the last 20 tests that were completed. PATIENT ID: Used to assign the patient identification number to a sample. Test results are then printed with the appropriate patient

Enters the information input via the keyboard into the microcomputer. DELETE TEST/CHEMISTRY SELECT: Delete Test: (Press SHIFT key.) Used to delete a test after a slide has been entered, identified and spotted. When a test is deleted, the slide must continue through the analyzer, DTE II Module, or DTSC II Module. The printout indicates that the test was deleted. Do not remove a slide manually once deleted. Chemistry Select: Used to identify a test if the analyzer is unable to read the information contained in the barcode. Press the CHEMISTRY SERVICE MODE/CAL MODE: Service Mode: (Press SHIFT key.) Used to enter and exit the service mode and to access various equipment options. Cal Mode: Used to enter and exit the calibration mode. Signals to the microcomputer that the analyzer is about to be calibrated and registers the PRINT: Advances paper in the printer when pressed during normal analyzer operation.

1-2

Operator's Manual VITROS DT II System

5/95. Reprinted 1/99.

1.2.2 Display Panel


The liquid crystal display panel is located above the keyboard and displays messages of up to two 40-character lines. In general, the first line of the display message tells you the status of the function you are in the process of performing, and the second line provides instructions on what to do next.

1.2.3 Printer and Results Printout


The printer is located to the left of the keyboard and is used to print out test results and other information. It provides a paper record.

1.3 Principles of Operation: VITROS DT60 II Chemistry System 1.3.1 Slide Identification
Place the VITROS DT Slide into the loading station of the analyzer. After loading a slide for the DT60 II System into the loading station, push the slide advance lever steadily and smoothly to move the slide into the spotting station. As the slide enters the spotting station, a bar code reader reads the laser bar code on the slide to identify the test and the slide generation number for the microcomputer. The test identification then appears on the display panel.

1.3.2 Slide Spotting


When the display says SPOT SLIDE WITH FLUID, insert the VITROS DT Pipette into the pipette locator and press down on the pipette button to dispense 10 uL of sample fluid onto the slide in the spotting station. An optical drop detector starts the analyzer's built-in timer and verifies that the slide has been spotted. Then, the analyzer automatically transports the slide into the incubator.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

1-3

1.3.3 Incubation and Readout


The incubator can hold a maximum of six slides stacked one on top of the other. Each slide enters the incubator at the top of the stack and exits from the bottom. The slides are incubated for approximately 5 minutes at 37C (98.6F). At the completion of the incubation period, the lower rack assembly automatically moves the slide from the bottom of the slide stack in the incubator to the read station. White and black reference readings are automatically taken. These readings, together with the slide reading, are used to determine the density of the color formed by the chemical reactions in the slide. As the lower rack assembly is retracted, the slide falls on top of the fiber-optics reflection system (FORS) head in the read station. The FORS head contains red, green, and yellow light-emitting diodes (LEDs). The appropriate LED is energized depending upon the type of slide present in the read station. The light is transmitted to the slide by fiber optics, and the light reflected off the slide is conducted to a photodetector within the FORS head. The microcomputer uses this reflectance reading, together with the white and black reference readings, a calibration model, and the stored calibration parameters, to calculate the concentration of the sample. This concentration is printed out by the analyzer's printer. The slide remains in the read station until the next slide pushes it into the slide disposal box at the rear of the analyzer.

1-4

Operator's Manual VITROS DT II System

5/95. Reprinted 1/99.

1.4 Principles of Operation: VITROS DTE II Module 1.4.1 Slide Identification


Place the DT Slide in the loading station on the moduleand use the slide advance lever to move the slide to the spotting station. As the slide enters the spotting station, a bar code reader in the module identifies the test and the slide generation number by reading the bar code on the slide. The test identification then appears on the analyzer's display panel.

1.4.2 Slide Spotting


The VITROS DTE Pipette, a dual pipette designed for use with the DTE II Module, allows you to simultaneously aspirate sample fluid and electrolyte reference fluid from the VITROS DTE Dual-Sample Cup. After the fluid is aspirated, you position the pipette over the slide in the spotting station. When you press down on the pipette button to dispense the fluids on the slide, a 3-minute timer is activated. When the slide is spotted, the indicator on the module lights up and begins flashing to alert you to wait before loading another slide.

1.4.3 Incubation and Readout


The electrometer assembly moves over the slide. The slide is then incubated in place for 3 minutes at 25C (77F). The electrometer has two contact points: one penetrates the contact area on the half of the slide where the reference fluid was dispensed, the other penetrates the contact area on the half where the sample fluid was dispensed. At the completion of the 3-minute incubation period, a tone sounds and the electrometer measures the potential difference between the two ion-selective electrodes in the slide. This potential difference is related to the difference in ionic concentration between the reference fluid and the sample fluid. This reading is used by the analyzer's microcomputer, together with the calibration parameters, to calculate the concentration of the electrolyte in the sample. The slide remains in this position until the next slide pushes it into the slide disposal box in the left side of the module.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

1-5

1.5 Principles of Operation VITROS DTSC II Module 1.5.1 Slide Identification


Place the DT Slide for the DTSC II into the pick-up station of the DTSC II Module. After the slide enters the slide pickup station, it is automatically moved to the bar code reader in the module. The bar code scanner identifies the test and the slide generation number on the slide. This information is then transmitted to the DT60 II System, where the test identification is displayed on its display panel.

1.5.2 Slide Spotting


The DT Pipette is designed for use with the DTSC II Module. A green indicator light on the DTSC II Module, and a message on the DT60 II System, signal you that you may spot the slide. You then insert the pipette into the pipette locator and press down on the pipette button to dispense 10 uL of sample fluid onto the slide in the spotting station. The spotting of the slide is automatically detected by an optical drop detector and the slide is transported to the preheat station.

1.5.3 Incubation and Readout


The slide is transported to the preheat station and then to the incubator where it is incubated in place at approximately 37C (98.6F) for a time specified by the tes-t. After incubation, while the slide is being transported to the read station, it momentarily stops to allow the system to take a white reference reading, a dark reference reading, and a voltage reference reading, and then transmits these readings to the DT60 II System for later use. At the completion of the reference readings, the slide is transported to the reflectometer read station where it is illuminated with light from a xenon flash lamp. At this point several readings are taken depending on the test. The light reflected from the analysis slide passes through an interference filter of the proper wavelength and onto a silicon photodetector. The electrical signals from the photodetector are fed to an analog-to-digital converter and the resulting data is sent to the DT60 II System. The microcomputer uses the reflectance readings, reference readings, and the calibration parameters to determine the concentration of the sample. When the slide readings are completed, the slide is automatically moved to the disposal station and into the slide disposal box.

1-6

Operator's Manual VITROS DT II System

5/95. Reprinted 1/99.

1.6 Supplies and Supply Handling and Storage 1.6.1 Storage Temperature Requirements
VITROS slide and fluid boxes are color coded in order to indicate the storage temperature of the slides and fluids. Light blue indicates the product must be frozen at temperatures less than -18CorOF. Purple indicates storage in refrigerator or freezer (<8C or 46F). Yellow indicates storage in a refrigerator at temperatures between 2 an.d 8C or 36 and 46F.

1.6.2 VITROS DT Slides


The DT60 II System, DTE II Module, and DTSC II Module each use a different type of DT Slide for analysis. The slides are individually wrapped for their protection during storage. The slide wrapper identifies the test and the slide lot number. The laser bar code on the slide is used by the analyzer and the associated modules to identify the slide and the generation number.

1.6.3 VITROS DT and DTE Pipettes and Micro Tips


The DT Pipette (a battery operated pipette) is supplied for use with your DT60 II System and DTSC II Module. The DTE Pipette is supplied with the DTE II Module. It is a dual sample pipette designed to permit you to simultaneously aspirate and dispense sample fluid and VITROS DT Reference Fluid. Disposable VITROS Micro Tips are used with these pipettes. These micro tips are designed to meet the specific metering requirements of the VITROS DT II System. A tip is used once to spot a single slide and is then discarded.

1.6.4 VITROS DT Calibrator Kits


Calibration of the analyzer requires the use of calibrator fluids which are provided in sets. Storage and stability requirements of calibrators are provided in the instruction sheet packaged with the fluids. The instruction sheet should be read prior to using the calibrators. Refer to the specific test Instructions for Use for special calibration precautions.

Rev. 2003-10-01

Operator's Manual VITROS DT II System

1-7

1.6.5 VITROS DT Control I and DT Control II


VITROS DT Control I and DT Control II contain known amounts of the same analytes that the analyzer measures in actual patient samples. Use these to compare analyzer results with known values which are provided with the control fluid. Quality control verifies the success of a calibration and allows you to monitor the long-term performance of the equipment. Refer to the specific test Instructions for Use for information on DT Controls I and II. Storage and stability requirements of unreconstituted and reconstituted fluids are provided in the instruction sheet packaged with the fluids and should be read prior to using controls. For specific analyte stability information, handling precautions and assay control values, refer to the assay sheet.

1.6.6 VITROS DT Reference Fluid


DT Reference Fluid is required for use with slides that are tested on the DTE II Module. Reference fluid is supplied in 10 mL squeeze bottles with a dropper-like tip that permits dispensing fluid into the DTE Dual-Sample Cup. The fluid is colored to permit you to easily view the amount aspirated. The bottle label identifies the fluid generation number, the lot number, and the expiration date. Storage and stability requirements are provided in the instruction sheet packaged with fluids and should be read prior to using reference fluids.

1.6.7 VITROS DTE Dual-Sample Cups


Single-use, disposable dual-sample cups are used in the sample holder of the DTE II Module. The sample cups contain two wells. As indicated on the sample carrier in the module, the sample to be analyzed is placed in the large well, and DT Reference Fluid is placed in the small well. The wells are positioned in the cup so that the DTE Pipette will aspirate both fluids simultaneously.

1.6.8 Printer Paper


Thermal printer paper is used in the analyzer's data printer. The paper is supplied in rolls 57 mm (2.25-inches) wide and 44mm (1.75-inches) in diameter.

1-8

Operator's Manual VITROS DT II System

Rev. 2003-10-01

1.6.9 VITROS DT Accessory Kit51


The VITROS DT Accessory Kit is supplied with each new system. The kit contains: One 3 mL pipette One package of 75 pipette tips for the 3 mL pipette One package of 100 2 mL plastic sample cups One package of 100 plastic sample cup caps One package of 100 transfer pipettes One box microwipes One package of 250 ViTROS Micro Tips One package of 1 60 VITROS DTE Dual-Sample Cups One VITROS DT Pipette battery recharger U.S. customers only

Rev. 2003-10-01

Operator's Manual VITROS DT II System

1-c

1 -10

Operator's Manual VITROS DT II System

5/95. Reprinted 1/99.

Operating Instructions
The previous section gave you an overview of your equipment's operating features. By following the operating instructions and illustrations in this section, you will discover how easy it is to make your VITROS DT60 II Chemistry System work for you. As you become more familiar with your DT60 II System, many of the frequently performed operating procedures explained in this section will become second nature.

2.1 Start-up Procedure


STEP 1. Prepare materials for use. ACTION TO TAKE

Allow the controls, slides, and reference fluids to come to room temperature (15-30 minutes). Unless otherwise noted on the control sheet packed with the control, reconstitute a new vial after 7 days. Write the date on the control and reference fluid when first opened. Allow patient samples to come to room temperature if refrigerated or frozen. Mix all fluids before using (gently invert several times).

2. Check power cord connections.

Check that all power cords are securely plugged into the proper outlets.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

2-1

This page has been left blank intentionally.

2-2

Operator's Manual VITROS 'I System

5/95. Repr'

1/99. <

2.1 Start-up Procedure (continued)


STEP 3. Turn DT60 II System on. ACTION TO TAKE The On/Off switch is on the back of the analyzer. Requires a 20-minute warm-up period. 4. Turn DTSC II Module on. The On/Off switch is on the back of the module. Requires a 5-minute warm-up period. Turning the DTSC II Module off and on every 24 hours will allow it to adjust for changes in lamp output. 5. Do daily maintenance. Empty all slide disposal boxes. Check the end of the.pipettes for residue and clean if necessary. See the Cleaning the DT Pipettes section of this Operator's Manual 6. Enter date. Press SHIFT and SERVICE keys. Key in option 1 7 and ENTER. Key in month, day, and year (NN-NN-NN) and ENTER. Press SHIFT and SERVICE to exit. 7. Run control materials. Run the controls on those tests that you will be using that day. See the How to Perform a Quality Control Test section of this manual.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

2-3

This page has been left blank intentionally.

2-4

Operator's Manual VITROS M System

5/95. Repr

* 1/99.

2.2 Testing Procedure 2.2.1 Pipetting Techniques


Samples to be analyzed require accurate fluid dispensing through pipetting. The importance of accurate pipetting procedures cannot be over-emphasized. Before attempting to analyze patient samples or performing routine calibration procedures, you should become thoroughly familiar with the pipetting procedures recommended for the DT Pipette and the DTE Pipette. Follow the simple instructions outlined on the next page. Collect and handle the patient sample according to standard laboratory procedures, centrifuging it to separate the serum or plasma. Place a cap on the container to avoid sample evaporation and contamination. Refer to Instructions for Use for proper storage of samples if analysis cannot be performed the same day that the sample is drawn.

Rev. 2003-10-01

Operator's Manual VITROS DT II System

2-5

This page has been left blank intentionally.

2-6

Operator's Manual VITROS M System

5/95. Repr

' 1/99.

2.2.1 Pipetting Techniques (continued)


COMMENT OR ACTION TO TAKE STEPS 1. Place disposable tips in the holder. 2. Check pipette(s). 2a. If the message I* R5P] appears in the display proceed to step 3. IMPORTANT: When the message appears in the display the DT P pette may be used to aspirate samples as long as the display indicates the presence of at least one aspiration |* Lo (]. The DT Pipette should remain plugged into the charger until it is fully charged. 2b. Plug the recharge unit into a wall outlet. NOTE: The DT Pipette only charges when the Long Term Storage switch is in the On position. 2c. Plug the power cord of the recharge unit into the power socket on the top of the DT Pipette. DT Pipette DTE Pipette

Power Socket

Rev. 2003-10-01

Operator's Manual VITROS DT II System

2-7

This page has been left blank intentionally.

2-8

Operator's Manual VITROS 'I System

5/95. Repr'

' 1/99.

2.2.1 Pipetting Techniques (continued)


COMMENT OR ACTION TO TAKE STEPS DT Pipette DTE Pipette

2d. Make sure the message \jh . [ appears in the display. If it does not, check the power cord and make sure it is attached 2e. The message Lo will appear in the display when the DT Pipette is ready to aspirate a 10 uL drop of fluid.
3. Insert tip(s).

Attach a disposable tip by pressing the tip cone of the DT Pipette into one of the tips in the tip holder. The tip will click into place when it is seated correctly.

To attach disposable tips, press the DTE Pipette firmly into two of the tips in the holder. The tips will click into place when seated correctly. Visually inspect tips to make sure that the tips project equally from the pipette.

\j
'!

y
2-9

//7

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

This page has been left blank intentionally.

2-10

Operator's Manual . VITROS 'I System

5/95. Repr

< 1/99.

2.2.1 Pipetting Techniques (continued)


COMMENT OR ACTION TO TAKE STEPS DT Pipette DTE Pipette

4. Insert and fill dual-sample cup.

Swing the sample holder into the loading position. Place a dual-sample cup into the holder so that the small well fits into the small depression in the holder and the large well fits into the large depression in the holder.

Gently invert the bottle of VITROS DT Reference Fluid to mix the fluid, and then squeeze at least 4 drops into the small well of the cup. Using a transfer pipette, pipette 4 drops (need at least 50 uL) of sample to be analyzed into the large well of the cup. IMPORTANT: Avoid bubbles in the fluid. Gently swing the sample holder back into place.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

2-11

This page has been left blank intentionally.

2-12

Operator's Manual VITROS M System

5/95. Repr'

' 1/99.

2.2.1 Pipetting Techniques (continued)


COMMENT OR ACTION TO TAKE STEPS 5. Aspirate fluids. DT Pipette Hold pipette in vertical position. Insert the tip below the surface of the fluid, but not to the bottom of the fluid container. Press the Start button to aspirate 10 uL of fluid. NOTE: The start button must be pressed twice to aspirate a sampje just before the messages f Ltffl] and f LoG] appear. This is an indication that the pipette needs to be charged. Remove the DT Pipette from the fluid within two seconds. IMPORTANT: A dotted line will appear in the display for two seconds while the DT Pipette is waiting. The pipette must be removed from the fluid before the dotted line disappears to prevent accidental aspiration of additional fluid. After the two second pause, the DT Pipette will aspirate 2 uL of air. Upon completion of the 2 uL pullback, the message j< d5P| will appear in the display. DTE Pipette Hold pipette in vertical position. Depress button and continue to hold button down as you insert pipette into pipette locator at the aspiration station. Slowly release pipette button and wait one second before removing pipette from the pipette locator in the aspiration station. Hold the pipette vertically whenever there is fluid in the.tips. If you tilt the pipette or lay it on its side, fluid might enter the mechanism, causing it to become clogged. If this occurs, clean the pipette immediately.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

2-13

This page has been left blank intentionally.

2-14

Operator's Manual VITROS II System

5/95. Repr

d 1/99.

2.2.1 Pipetting Techniques (continued)


COMMENT OR ACTION TO TAKE STEPS 6. Remove excess fluid. DT Pipette Remove any droplets which may be clinging to the outside of the tip by taking a laboratory tissue and wiping the outside of the tip in a light, quick, downward motion. Do not wipe across the bottom of the tip. IMPORTANT: If the tip is not wiped, test results may not be accurate. 7. Check fluid volume. Visually check the fluid level inside the tip. If fluid volume is not as shown, dispense sample, eject tip, and repeat steps 1-6. DTE Pipette To remove any droplets which may be clinging to the outside of the tips, take a laboratory tissue and wipe the outside of the tips in a light, quick, downward motion. Do not wipe across the bottom of the tip. If the tips are not wiped, test results may be inaccurate. Visually check fluid level in both tips as shown in the illustration. Both tips should have approximately equal fluid levels. If fluid level is not as shown, dispense sample, eject tips, and repeat steps 1 -6. NOTE: Inadequate fluid in tip(s) can lead to errors in test results.

Fluid Level Check for Air Space at Tip


NOTE: Inadequate fluid in tip can lead to errors in test results.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

2-15

This page has been left blank intentionally.

2-16

Operator's Manual VITROS ' II System

5/95. Repr

i 1/99.

2.2.1 Pipetting Techniques (continued)


COMMENT OR ACTION TO TAKE STEPS DT Pipette For procedures on slide spotting refer to Section 2.2.2. Visually check the tip to assure that the fluid was completely dispensed from the tip. If fluid is still visible, dispense the fluid into a tissue and repeat the test with a fresh tip. IMPORTANT: Never eject a tip that has fluid in it. Always dispense the fluid completely before ejecting the tip. Press the tip eject lever to eject the used tip. DTE Pipette For procedures on slide spotting refer to Section 2.2.2. Check that the fluid was completely dispensed from the tips. If fluid is still visible, press the button to dispense the fluid into a tissue. Press the latch button to eject the used tips. Dispose of tips. NOTE: Fluid should not be used if it has been in the dual-sample cup for more than 5 minutes. If so, take a clean dual-sample cup and pipette fresh fluid.

8. Spot the slide. 9. Check pipette tips.

1O.Ejecttip(s)

Dispose of tip. NOTE: Use tips only once. Always use a new tip for each aspiration, even if it is drawn from the same fluid.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

2-17

This page has been left blank intentionally.

2-18

Operator's Manual VITROS 'I System

5/95. Repr'

' 1/99.

2.2.2 Steps for Analysis on the VITROS DT II System


COMMENT OR ACTION TO TAKE STEPS 1. Warm slides to room temperature. 2. Load slides. GENERAL INFORMATION This takes approximately 15 minutes DT60 II System DTSC II Module DTE II Module

Position the slide into the loading station. IMPORTANT: Inserting or removing a slide without following the analyzer prompts can cause unexpected results: Wait until the "ANALYZER READY" message appears before inserting a slide. DO NOT remove slide from spotting station after insertion unless directed by the analyzer.

Manually push the slide advance lever to move slide into spotting station.

Manually insert the slide. It is automatically carried to the spotting station.

Manually push the slide advance lever to move slide into the spotting station.

c m
0 O j

See illustrations for proper insertion of slide.

Notch in first Bar code up

Notch to right Bar code up

Notch in first Bar code down

Rev. 2004-03-30

Operator's Manual VITROS DT II System

2-19

This page has been left blank intentionally.

2-20

Operator's Manual VITROS 'I System

5/95. Repr

' 1/99.

2.2.2 Steps for Analysis on the VITROS DT II System (continued)


COMMENT OR ACTION TO TAKE STEPS 3. Enter patient identification (optional). GENERAL INFORMATION Analyzer will display the message (optional) to enter information with "1D=". To enter any patient identification number up to 10 characters, press the PATIENT ID key and the numbers or characters you wish to enter, then press the ENTER key. If you do not wish an ID to printout, press the PATIENT ID and CLEAR key and then press the ENTER key. 4. Insert tips onto the DT Pipette or DTE Pipette. 5. Aspirate the fluid. Press tips firmly into place. Use 1 new tip. Use 1 new tip. Use 2 new tips. DT60 II System DTSC II Module DTE II Module

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

2-21

This page has been left blank intentionally.

2-22

Operator's Manual VITROS 'I System

5/95. Repr

'1/99.

2.2.2 Steps for Analysis on the VITROS DT II System (continued)


COMMENT OR ACTION TO TAKE STEPS GENERAL INFORMATION DT60 II System DTSC II Module DTE II Module

6. Spot the slide.

Gently insert pipette(s) into locator. The pipette must be fully seated in the pipette locator. Press the Start button to dispense 10 uL of fluid onto the slide. A dotted line will appear in the display after which the message (* R5P] or the message la will appear in the display indicating that the DT Pipette is ready for the next aspiration/ dispense cycle. IMPORTANT: DO NOT remove slide from spotting station after insertion unless directed by the analyzer.

An audible tone indicates that the slide has been spotted. Remove the DT pipette from the pipette locator within one second of the tone.

Green flashing light indicates "ready-tospot" status. At the sound of the tone promptly remove the pipette from the locator. Delay in the removal of the pipette may result in inaccuracy and imprecision.

Depress the pipette button and continue to hold it depressed. With the button still depressed, promptly but slowly, remove the pipette from the locator. An audible tone will tell you when the slide has been spotted. IMPORTANT: Do not release the pipette button until you have removed it from the pipette locator. Reaspiration of fluid that has already been dispensed on the slide may occur. Red indicator light will flash while slide is incubating.

7. Eject tip(s).

Always use a new tip(s) for each sample analyzed.

Dispose of tip.

Dispose of tip.

Dispose of tips.

Rev. 2004-03-30

Operator's Manual VITROS DT II System

2-23

This page has been left blank intentionally.

2-24

Operator's Manual VITROS II System

5/95. Repr

1 1/99.

2.2.2 Steps for Analysis on the VITROS DT II System (continued)


COMMENT OR ACTION TO TAKE STEPS 8. Repeat for other tests. GENERAL INFORMATION Repeat steps 1 -7 for additional sample analysis once analyzer is ready to accept another slide. DT60 II System Display will read ANALYZER READY. DTSC II Module Display will read DTSC MODULE READY. DTE II Module Display will read DTE MODULE READY.

IMPORTANT TIPS
Allow the slides to reach room temperature before use. Press tip(s) firmly onto pipette. Check fluid level in tip(s). Wipe excess fluid from Ihe oulside of tips. Handle pipette carefully while aspirating fluid and spotting slide. Orient slides as shown in illustrations. Enter the patient ID when prompted by the analyzer. If an ID is nol enlered, [he previous ID that was entered will print out. When DT60 il System is turned on, the number 1 is an analyzer-assigned ID number for your first sample until the operator changes it. Always follow the analyzer prompts. Wait until the "ANALYZER READY" message appears before inserting a slide.

Rev. 2004-03-30

Operator's Manual VITROS DT II System

2-25

This page has been left blank intentionally.

2-26

Operator's Manual VITROS M System

5/95. Repr

< 1/99.

2.3 Calibration Data Module and Chemistry Language Module 2.3.1 Description of Calibration Data Module
The calibration data module (CDM) is an electronic device contained in a protective carrier. The CDM contains several different types of data required to perform various tests on the DT60 II System and the DTE II and DTSC II Modules (for example, the generation number and the calibration kit number). OCD will issue a new CDM when: 1. You receive slides with a new generation number that is not supported in your current CDM. (The generation number appears on the individual slide wrapper and slide cartons, and on the bottle labels for Reference Fluid to define a formulation and process of manufacture.)

2. You receive calibrators having a new kit number which is not supported in your current CDM. The CDM must be installed before you use the new calibrators in order for the instrument to use the correct calibration data.
CAUTION: A CDM is a delicate electronic device. Handle with care.

2.3.2 Description of Calibration Language Module


Like the calibration data module, the chemistry language module (CLM) is an electronic device contained in a protective carrier. The CLM contains data on the messages that appear on the screen display and information needed to perform all tests available on the analyzer. Like the CDM, the CLM is designed to plug into back of the analyzer. CAUTION: The CLM is a delicate electronic device. Damage can occur. Handle with care. A new CLM is provided whenever an additional test becomes available unless your current CLM already contains the test. French, German, and Italian language CLMs are available to non-U. S. customers upon request.

Rev. 2003-10-01

Operator's Manual VITROS DT II System

2-27

This page has been left blank intentionally.

2-28

Operator's Manual VITROS " " II System

5/95. Repr"

' 1/99.

2.3.3 How to Install a New Calibration Data Module (CDM) and a Chemistry Language Module (CLM)
Follow the steps outlined below for simple installation instructions for both the CDM and CLM.
STEP 1. Turn the analyzer off. 2. Remove the old C D M or CLM. ACTION TO TAKE

The analyzer must be turned off before installing either a new CDM or CLM. Begin removing the old CDM or CLM by opening the door on the back of the analyzer to gain access to the compartment that houses the CLM and CDM. Read the labeling on the side of the compartment to make sure that you are unplugging the correct component. Unplug the old CLM or CDM as shown in the illustration.

5/95. Reprinted 1/99.

Operator's Manual . VITROS DT II System

2-29

This page has been left blank intentionally.

2-30

Operator's Manual VITROS I! System

5/95. Repr

\ 1/99.

2.3.3 How to Install a New Calibration Data Module (CDM) and a Chemistry Language Module (CLM) (Continued)
STEP 3. Install the new CLM or C D M . ACTION TO TAKE Hold either component by the side edges and carefully remove it from its packaging. Position the new CLM or CDM as shown on the body of the analyzer. Note the positioning of the tabs. Check the alignment of the carrier to the receptacle before inserting the CLM or CDM. Check the labeling to make sure that you have inserted the correct component into the correct receptacle. For CLM installation: Calibrate the analyzer for the new test if you received the CLM with the materials for the test. If you changed the CLM only to change the language on the display screen then there is no need to recalibrate. NOTE: If the print is jumbled on the printer, the CLM and CDM may be in the wrong positions.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

2-31

This page has been left blank intentionally.

2-32

Operator's Manual . VITROS 'I System

5/95. Repr

' 1/99.

2.4 Normal Shutdown Procedure


The analyzer can be left on at the end of each day, however, the DTSC II Module should be turned off at the end of each day. Turning the analyzer off results in the loss of tests that may remain in the incubator and test results that have not been printed. Turning the analyzer off has no effect on the instrument's memory, and does not result in loss of calibration values or loss of other information stored in the memory. Do not have the DTSC II Module on if the DT60 II System is turned off. When turning the analyzer off, follow the steps outlined here. 1. Check incubator for tests in progress. First, check that there are no unread slides in the analyzer through use of the TEST IN PROGRESS/TEST COMPLETE key. If the analyzer is connected to a VITROS DTE II Module, check the red indicator on the module. If the indicator is flashing, wait until the test result is reported and the indicator stops flashing. 2. Turn analyzer off. When you are sure that all tests are complete, turn off the DTSC II Module. Then turn the DT60 II System off by moving the main power switch to the OFF position.

2.5 Emergency Shutdown


In case of an emergency, simply turn the main power switch on the DTSC II Module to OFF, then turn off the DT60 II System. When you turn the analyzer on again, you will have to repeat the analysis for all tests that remained in the incubator at the time of the shutdown.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

2-33

This page has been left blank intentionally.

2-34 \

Operator's Manual '("System

5/95. Repr

' 1/99.

Coronary Risk Classification (CRC)


The analyzer's microcomputer has the ability to perform Coronary Risk Classification (CRC) analysis for the purpose of determining five- and ten-year coronary risk probabilities. The American Heart Association (AHA) recognizes this analysis which is based on an update of the coronary risk assessment methodology of the Framingham Heart Study conducted by the National Heart, Lung and Blood Institute (Anderson, K. M., Wilson, P. W. R, Odell, P. M., and Kannell, W. B., "An Updated Coronary Risk Profile," Circulation: Vol. 83, No. 1Jan. 1991). This section provides you with instructions for accessing the CRC program, entering the appropriate data needed to calculate coronary risk probabilities, and printing the five- and ten-year risk probabilities.

3.1 CRC Overview


To develop a coronary risk profile, the AHA recognizes various risk factors including sex, age, cigarette smoking, high blood pressure, high levels of serum cholesterol, low HDL-cholesterol, diabetes and ECG abnormalities. Although these are not the only accepted risk factors, they represent information which is easily available from a doctor's office and is without patient discomfort. Values assigned for each risk factor are awarded points, based on the relative degree of risk. The sum of all points then determines the CRC fiveand ten-year risk probabilities.
CRC five- and ten-year risk probabilities take into account eight risk factors and should be used as a guide

only. The test does not take into account other risk factors, e.g., obesity, x-ray cardiac enlargement, lack of physical activity, heredity etc. Furthermore, the probabilities represent average values over a given population and not necessarily the experience of any one person. Coronary Risk Classification should be used with other information and parameters available from clinical patient evaluation. The CRC predictions provide a diagnostic tool, but are not a substitute for the physician's judgment.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

3-1

This page has been left blank intentionally.

3-2

Operator's Manual VITROS "" '1 System

5/95. Repri

' 1/99.

The CRC program allows the user to input "what if?" values for a given profile in order to determine the overall effect of that change on the five- and ten-year risk probability. For example, a 55-year-old male smoker who has a five-year CRC of 22% may see that he can reduce his risk probability to 1 6% if he gives up smoking. Of course, there is no assurance that lowering any given risk factor will in reality reduce overall risk. Clinical trials of risk-factor alteration, however, provide positive evidence that improving the risk-factor profile will lower the risk of coronary disease.

3.2 Entering Data for Coronary Risk Classification


1 . Access the CRC option Press SHIFT, then SERVICE/CAL MODE. When the display prompts you to ENTER OPTION NO., key in option 24 and press ENTER to create, enter and modify CRC records. You may only enter this option if there are no samples processing. If samples are processing, the analyzer will beep and display WAIT - TESTS IN PROGRESS until it completes the test. 2. Enter Patient ID At the CRC ENTER PATIENT ID prompt, the current CRC record in memory will be displayed if one exists. You have three choices: Press ENTER if you wish to use the patient ID currently displayed. Key in up to 1 0 characters and press ENTER if you wish to enter a new patient ID. You cannot change the patient ID, once it has been entered. Press the PATIENT ID key if you wish to use a patient ID in memory other than the patient ID currently displayed.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

3-3

This page has been left blank intentionally.

3-4

Operator's Manual . VITROS 'I System

5/95. Repr

' 1/99.

3.2 Entering Data for Coronary Risk Classification (continued)


The CRC program allows you to maintain CRC records for up to three different patient IDs at one time. If a fourth record is created, it replaces the oldest of the three existing CRC records. When you attempt to create a fourth record, the analyzer will tell you if the oldest record has not been reported. CRCxxxxxxxxxNOT REPORTED DELETE TEST TO REPLACE It will identify the patient ID that will be replaced and will give you two options. 1. Replace the oldest record with the new patient ID by pressing SHIFT and DELETE TEST. 2. Return to the CRC ENTER PATIENT ID prompt with no replacement. To do this press ENTER. You may also print from the patient ID screen. Touch PRINT for a printout of the CRC report. The analyzer will indicate which, if any, data is missing. It will not generate a coronary risk classification until all information needed to calculate the test has been entered. 3. Enter Sex Key in 0 for female or 1 for male, as appropriate. Press ENTER. If you are working with an established record and do not wish to make a change, press ENTER to continue.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

3-5

This page has been left blank intentionally.

3-6

Operator's Manual VITROS 'I System

5/95. Repr"

'1/99.

3.2 Entering Data for Coronary Risk Classification (continued)


4. Enter Age Enter the current age of the patient and press ENTER. The program is based on an age range of 30 to 74 years. If an age is entered that is less than 30, the program will accept the value but will automatically set that age value to 30; similarly, if an age is entered that is greater than 74, the program will set that age value to 74. Any data modified by CRC program in this way will be noted with both a plus sign (+) next to the age and by the words "DATA SET TO AHA CHART LIMITS" on the printed CRC report. If you are working with an established record and do not wish to make a change, press ENTER to continue. 5. Enter Systolic Blood Pressure Enter the systolic blood pressure of the patient and press ENTER. If an SBP value is less than 98 or greater than 185, the program will accept the value but will automatically set that value to 98 or 185, respectively. When the SBP field is modified this way, a plus sign (+) and the words "DATA SET TO AHA CHART LIMITS" will appear on the printed CRC report. SBP measurements should be taken with the patient seated. The average of at least two readings is preferred. If you are working with an established record and do not wish to make a change press ENTER to continue.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

3-7

This page has been left blank intentionally.

3-8

Operator's Manual VITROS "~~ II System

5/95. Repr

^ 1/99.

3.2 Entering Data for Coronary Risk Classification (continued)


6. Enter Smoker or Nonsmoker Enter 0 if the patient is not a smoker and has not smoked for the past 12 months. Enter 1 if the patient is a smoker or has quit smoking within the past 12 months. Once your selection is made, press ENTER. NOTE: The program is based on data which does not address the "degree" of smoking, i.e., heavy smokers and light smokers. A distinction is made only between smokers and nonsmokers. If you are working with an established record and do not wish to make a change, press ENTER to continue. 7. Enter Diabetic or Nondiabetic Enter 0 for nondiabetic or 1 for diabetic, whichever is appropriate, and press ENTER. For purposes of coronary risk assessment, the program defines a diabetic patient as one who is currently under treatment with insulin or oral hypoglycemic agents OR who has a fasting glucose level of 140 mg/dL or greater. If you are working with an established record and do not wish to make a change, press ENTER to continue. 8. Enter ECG-LVH Enter 0 if the patient tests negative for left-ventricular hypertrophy by ECG. Press 1 if the patient tests positive. Press ENTER to continue. LVH-ECG consists of finding tall R-waves in leads reflecting potentials from the left ventricle. The R-waves are accompanied by nonspecific S-T or T-wave abnormalities. Enter 0 for ECG-LVH to get a rough estimate of a person's coronary risk classification, when ECG data are unavailable. It should be recognized, however, that in a small fraction of people the CRC assessment will be inappropriately low since ECG-LVH is a strong risk factor when it is reported as positive. See the CRC Supplement for further information. If you are working with an established record and do not wish to make a change, press ENTER to continue.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

3-9

This page has been left blank intentionally.

3-10

Operator's Manual VITROS "I System

5/95. Repr

'1/99.

3.2 Entering Data for Coronary Risk Classification (continued)


9. Enter CHOL value

There are 3 approaches to entering the CHOL and HDLC values. 1. You may run up to 10 CHOL tests and 10 HDLC tests (the analyzer has the capacity to remember the results for 20 tests) and then enter the corresponding CRC data for those 10 patients. After a complete set of data is stored for a patient, you may successfully request a printout of the Coronary Risk Classification results. 2. You may enter the CRC data for up to three patients at a time, then run the CHOL and HDLC tests. The analyzer will automatically printout the CRC results after printing the results for the chemistry tests. 3. You may run the CHOL and HDLC tests and keep a record of the test results for input into the CRC option at a later time. This allows you to run the CRC tests at your convenience and not worry about the 20result capacity in the buffer. Assuming the CHOL test was run on the patient sample previous to entering CRC data, the cholesterol value will automatically be displayed on the analyzer screen. If the screen is blank, the CHOL value needs to be entered. Once the data has been computed and printed, you may enter a new CHOL value replacing the current value or alter other risk parameters, permitting you to conduct "what if?" tests. NOTE: Values that printout GREATER THAN ANALYZER RANGE will not be included in the 20 report buffer. Once the sample has been diluted it will not be considered multiple. The analyzer will automatically print a result based on the diluted sample. The CHOL value must be multiplied by two and entered into the data to replace the current (diluted) value. You will receive an incorrect CRC result if the
data is not modified.

Both CHOL and HDLC require manual entry if either was multiple within the last 20 results. In this case, the word MULTIPLE will be displayed on the screen. Enter the appropriate CHOL value. The printed CRC report will be marked with an asterisk (*) next to the value indicating that the value was manually entered or "User Modified" and with a pound sign (#) indicating multiple results. Both CHOL and HDLC will be marked as multiple if either was multiple. If a CHOL value is less than 139 mg/dL or greater than 330 mg/dL, the program will accept the value but will automatically setthat value to 139 mg/dL or 330 mg/dL, respectively. When the CHOL field is modified in this manner, a plus sign (+) and the words "DATA SET TO AHA CHART LIMITS" will appear on the printed CRC report.

5/95. Reprinted 1/99.

Operator's Manual . VITROS DT II System

3-11

This page has been left blank intentionally.

3-12

Operator's Manual VITROS 'I System

5/95. Repr

' 1/99.

3.2 Entering Data for Coronary Risk Classification (continued)


10. Enter HDLC value As mentioned in step 9, there are 3 approaches to entering HDLC values. If the screen is blank, the HDLC needs to be run or the value entered manually. If an HDLC value is less than 25 mg/dL or greater than 96 mg/dL, the program automatically sets that value to 25 mg/dL or 96 mg/dL, respectively. When the HDLC field is modified in this manner an asterisk (+) and the words "DATA SET TO AHA CHART LIMITS" will appear on the printed CRC report. NOTE: If all other criteria are in the analyzer memory and the HDLC has been pre-diluted, the analyzer will automatically print a result based on the diluted sample. The HDLC value must be multiplied by two and entered into the data to replace the current (diluted) value. You will receive an incorrect CRC result if the data is not modified. If you tend to manually enter your results, make sure you multiply the diluted value by two when entering the number. If you enter the CRC data after running the tests, make sure you modify the HDLC data before requesting a CRC report.

3.3 Printing CRC Results


Press PRINT if you wish to have the keyed-in values computed and a coronary risk classification report printed. The printed report lists the CRC values you entered for each field along with the five- and ten-year risk factors and it prints the peer average for 10-year risk within the appropriate gender. If all CRC data is available but the computed CRC total point value exceeds the American Heart Association chart maximum of 32, the following message will be printed to finish the CRC printout: ************************ AHA CHART EXCEEDED **** ********** ********** Press ENTER if you wish to return to the PATIENT ID prompt to add missing data or modify data before printing. The program will be unable to calculate the five- and ten-year risk probability if any field of information is missing. Note the meaning of certain indicators that may precede entered data (see sample CRC reports on next two pages): Operator's Manual VITROS DT II System 3-13

5/95. Reprinted 1/99.

This page has been left blank intentionally.

3-14

Operator's Manual - VITROS 'I System

5/95. Repr

' 1/99.

3.3 Printing CRC Results (continued)


plus sign (+) DATA SET TO AHA CHART LIMITS. Indicates that the entered value was not within the limits set by the program. Subsequently, the program modified the entered value. asterisk (*) USER MODIFIED. Data was modified after the CRC report was printed, e.g. smoker vs. nonsmoker. It also indicates a CHOL or HDLC value was entered manually AND the manual entry was made prior to printing the CRC report. pound (#) MULTIPLE RESULTS. Indicates that multiple results were found for either CHOL and/or HDLC prior to printing the CRC report. Both are marked a multiple if either had multiple results prior to printing. 12. Press SHIFT, then SERVICE/CAL MODE to exit option 24. 13. Press SHIFT, then SERVICE/CAL MODE again to exit the service mode.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

3-15

This page has been left blank intentionally.

3-16

Operator's Manual VITROS " 'I System

5/95. Repr"

' 1/99.

3.3 Printing CRC Results (continued)


1. CHOL & HDLC report, followed by CRC prediction and user-modified CRC prediction. 2. Age and SBP were set to AHA chart limits
*"*** ** *%*~**Ni*y<***y I.D: 5 89-04-91 ******** *********** CHOL ******************* MODIFIED CRC ******************* PT ID 190 FEMALE 45 VEARS 113 MM HS SBP * NON-SMOKER NON-DIABETIC ECG-LVH NEGATIVE * CHOL 189 MG/DL HDLC 46 MG/DL * USER MODIFIED IS 2m<

******************* I.D: 99-04-91 ******************* CHOL

HDLC 45 MG/DL ******************* CRC ******************* PT ID = FEMALE + 74 VEARS + 185 MM HG SBP NON-SMOKER NON-DIABETIC ECG-L'JH NEGATIVE CHOL 163 MG/DL HDLC 45 MG/DL + DATA SET TO CHART LIMITS 5 VR RISK b'-i 19 VR RISK 1 ' . 3AVERAGE 10 VEAR RISK FOR FEMALE 76-74 VEARS: 12X ******************* *******************

HDLC 46 MG/DL ************** CRC


- ^ ^ ^ ^ - ^ V ^ V V V V ^ ^^ ^^ ^ ^p ^ ^ ^ ^ ^ ^

PT ID 190 FEMALE 45 VEARS 118 MM HQ SBP SMOKER NON-DIABETIC ECG-LVH NEGATIVE CHOL 239 MG/DL HDLC 5 VR RISK 46 MG/DL 3":

5 VR RISK 19 VR RISK

10 VR RISK 6": AVERAGE 19 VEAR RISK FOR FEMALE 45-49 VEARS: 5>. ******************* *******************

AUERAGE 10 VEAR RISK FOR FEMALE 45-49 VEARS: 5'< ******************* *******************

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

3-17

This page has been left blank intentionally.

3-18

Operator's Manual VITROf II System

5/95. Rep'

i 1/99.

3.3 Printing CRC Results (continued)


3. Multiple reports for CHOL and/or HDLC followed by CRC report with manual entry of CHOL and HDLC.
******************* CRC ******************* PT ID 19 FEMALE 37 VEARS 12@ MM HG SBP SMOKER NON-DIABETIC ECG-LVH NEGATIUE #CH0L MULTIPLE #HDLC MULTIPLE
# MULTIPLE RESULT MISSING CRC DATA ******************* ******************* *******************

4. CHOL and HDLC followed by CRC report indicating risk factors were greater than AHA Chart Limits. *******************
09-94-91 ******************* CHOL 231 ItQ/DL

5. CRC report requested without all


information needed to make prediction.

1.0:

15

HDLC 45 MQ/DL ******************* CRC ******************* PT ID It FEMALE 72 YEARS 131 MM HG SBP SMOKER DIABETIC ECS-LUH POSITIUE CHOL 231 MG/DL HDLC 45 MQ/DL ******************* AHA CHART EXCEEDED ******************* *******************

******************* CRC ******************* PT ID 2? MALE 48 VEARS 119 MM HG SBP NON-SMOKER DIABETIC ECG-LUH NEGATIUE CHOL MISSING HDLC MISSING MISSING CRC DATA ******************* *******************

MODIFIED CRC ******************* PT ID 10 FEMALE Z7 VEARS 120 MM HG SBP SMOKER NON-DIABETIC EC6-LUH NEGATIUE *#CH0L 174 MG/DL *#HDLC 59 M6/-DL * USER MODIFIED
# MULTIPLE RESULT 5 VR RISK

10 VR RISK <2'/. AVERAGE 10 VEAR RISK FOR FEMALE 35-39 VEARS: <i\ ******************* *******************

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

3-19

This page has been left blank intentionally.

3-20

Operator's Manual VITROS H T II System

5/95. Repn

-I 1/99.

Derived Tests
The VITROS DT II System has the ability to calculate derived tests based on the results of related chemistry components. This section describes how to access the options for derived tests, how to process derived tests, and how to print derived test results.

3.4 Overview
Derived test processing is available for: 1. Globulin (GLOB) and the Albumin-Globulin Ratio (A/G). Calculations are derived from ALB and TP test results. 2. Very low density lipoproteins (VLDL), low density lipoproteins (LDL) and the cholesterol/high density lipoprotein cholesterol ratio (CHOL/HDLC Ratio). These tests are collectively called the Lipid Calculation. Calculations are derived from CHOL, HDLC and TRIG test results. 3. Anion gap (AGP). This calculation is derived from Na + , Cl" and CO 2 test results. For derived test processing to occur, the following conditions must be met: The derived test must be turned on by selecting the appropriate Option number prior to running tests on the component chemistries. Ajj of the components required to calculate the derived test result must be within the last 20 tests reported. There must be no more than one test result stored in the analyzer for the component and patient ID just tested. No multiple results are allowed. IMPORTANT To assure completion and accuracy of derived test processing: Component chemistry values must not be based on a dilution factor. A 1-10 character patient ID must be entered for the component tests prior to spotting the necessary

slides with fluid.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

3-21

This page has been left blank intentionally.

3-22

Operator's Manual VITROS M System

5/95. Repr"

< 1/99.

3.5 GLOB and A/G


These derived tests are calculated from ALB and TP test results as follows: GLOB

A/C

TP-ALB ALB/GLOB

To access globulin and the albumin/globulin processing: Press SHIFT, then SERVICE/CAL MODE to enter the service mode. You may only enter this option if there are no samples processing. If samples are processing, the analyzer will beep and display - WAIT - TESTS IN PROGRESS until it completes all tests. When the display prompts you to ENTER OPTION NO., key in 25 and press ENTER. Press 1, then ENTER to turn on GLOB and A/G Ratio derived test processing. Press SHIFT, then SERVICE/CAL MODE again to exit the service mode. Now you are ready to run the component tests as outlined in section 2 of this manual, Operating Instructions. Once the patient ID is entered and both the ALB and TP slides are analyzed, GLOB and A/G test results will automatically be calculated and printed along with the ALB and TP test results. If more than one value for ALB or TP for a given patient ID appears in the last 20 results buffer, the test name and MULTIPLE RESULTS Will appear on the printout followed by the name of the derived test and NO RESULT-COMPUTE MANUALLY.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

3-23

This page has been left blank intentionally.

3-24

Operator's Manual VITROS 'I System

5/95. Repr

'1/99.

3.5 GLOB and A/G (continued)


To turn globulin and the albumin/globulin processing off: Press SHIFT, then SERVICE/CAL MODE. When the display prompts you to ENTER OPTION NO., key in 25 and press ENTER. Press 0 to turn off GLOB and A/G derived test processing. Press SHIFT, then SERVICE/CAL MODE again to exit the service mode.

1. Results from component tests, ALB and TP, followed by GLOB and A/G results.

2. A 1-10 character patient ID was not entered prior to running ALB or TP tests.

3. The analyzer cannot calculate results for GLOB or A/G if there are multiple results for either of the component tests.

******************* I.D: 5154 81-20-92 ******************* ALB 3.0 8/DL TP 4.5 G/DL ******************* I.D: 5i5d TP 4.5 G - D --L ALB 3.0 G/DL
GLOB 1 . 5 6/-DL A/G 2.0 *******************

61-28-92 * ft************** ALB Z.B

1,1

1
* : : . * * * * * * * * * * *

***

*.***,-,

8..: j-'DL

TP

TP
j,'DL

3.6
ALB

******************* I.D! TP BLANK PATIENT ID HQ RESULT COHPUTE HAHUALLV *******************

5. i I.D:
Tp

j/DL

* * * * x * : * * * * * * * * * * * *

HL'-'.PLE RESULTS
GLOB,A,G HO
****:/*:yt

RESULT

COMP.Ti MANUALLV
* * * * * * * * * * * *

Operator's Manual V1TROS DT II System

3-25

This page has been left blank intentionally.

3-26

Operator's Manual VITROS 'I System

5/95. Repr

' 1/99.

3.6 Lipid Calculation (VLDL, LDL and CHOL/HDLC Ratio)


These derived tests are calculated from TRIG, CHOL and HDLC test results as follows: VLDL = TRIG/5.0 TRIG/2.2 LDL CHOL/HDLC ratio = = CHOL-HDLC-VLDL CHOL/HDLC (Conventional units) (SI units)

To access VLDL, LDL and CHOL/HDL Ratio processing: Press SHIFT, then SERVICE/CAL MODE to enter the service mode. You may only enter this option if there are no samples processing. If samples are processing, the analyzer will beep and display - WAIT - TESTS IN PROGRESS until it completes all tests. When the display prompts you to ENTER OPTION NO., key in 26 and press ENTER. Press 1, then ENTER to turn on the lipid calculation derived test processing. Press SHIFT, then SERVICE/CAL MODE again to exit the service mode. Now you are ready to run the component tests as outlined in section 2 of this manual, Operating Instructions. Once the patient ID is entered and CHOL, HDLC and TRIG slides are analyzed, VLDL, LDL AND CHOL/ HDLC ratio results will automatically be calculated and printed along with the CHOL, HDLC, and TRIG test results. If more than one value for CHOL, HDLC or TRIG for a given patient ID appears in the last 20 results buffer, the test name and MULTIPLE RESULTS will appear on the printout followed by the name of the derived test and NO RESULT-COMPUTE MANUALLY.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

3-27

This page has been left blank intentionally.

3-28

Operator's Manual VITRO II System

5/95. Rep

d 1/99.

3.6 Lipid Calculation (VLDL, LDL and CHOL/HDLC Ratio) (continued)


VLDL and LDL will not be calculated if the TRIG value is greater than or equal to 400 mg/dL (4.52 mmol/L). In this case, the error message NO RESULT TRIG TOO HIGH will be printed. To turn the lipid calculation derived test processing off: Press SHIFT, then SERVICE/CAL MODE. When the display prompts you to ENTER OPTION NO., key in 26 and press ENTER. Press 0, then ENTER to turn off the lipid calculation derived test processing. Press SHIFT, then SERVICE/CAL MODE again to exit the service mode.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

3-29

This page has been left blank intentionally.

3-30

Operator's Manual VITROS M System

5/95. Repr

' 1/99.

3.6 Lipid Calculation (VLDL, LDL and CHOL/HDLC Ratio) (continued)


1. Results from component test, TRIG, followed by the VLDL result. Results from component tests, CHOL and HDLC, followed by LDL, VLDL and CHOL/HDLC results. 2. A 1-10 character patient ID was not entered prior to running CHOL, HDLC, or TRIG.
*******************

3. The analyzer cannot calculate results for LDL, VLDL, or CHOL/HDLC if there are multiple results for any of the component tests.
* *** * * *: * ********** f

I. o;
01-20-92 * * * * ******** * * * * * * * CHOL 160 CIG-'DL HDLC 66 riG.-'DL ******************* I.D: HDLC BLANK PATIENT ID Cv-H,LDL HO R E S U L T COMPUTE MANUALLV I.D: 2" 5 61-24--92 ******************* CHOL 232 MG.'DL CHOL

*******************
I.D: 78816 01-28-92 ******************* TRIQ 125 MG-'DL ******************* I.D: 78916 TRIG 125 MQ/DL ULDL 25 PfG/DL *******************

234
TRIG

Mi

255 MG/DL
******************* I.D: 25 TRIG 255 MS/DL ULDL 51 MG-'DL fc***************** ******************* HDLC 52 f1G-'0L ******************* I.D: 25 CHOL MULTIPLE RESULTS
C / H , L D L HO RESULT COMPUTE MANIJALLV *******************

*******************
CHOL

**************+***
TRIG 128 TIQ/OL ******************* I. D : TRIG BLANK PATIENT ID ULDL,LDL NO RESULT COMPUTE MANUALLV *******************

163 MQ/DL
HDLC

65 MG.'DL ******************* I.D: 70816


TRIG CHOL HDLC 125 MS/DL 163 MG/DL 65 WG/DL

LDL 73 MQ^DL ULDL 25 MG/DL CHOL-'HDL RATIO 2.5 *******************

5/95. Reprinted 1/99.

Operator's Manual VITROS DT I! System

3-31

This page has been left blank intentionally.

3-32

Operator's Manual VITROS '! System

5/95. Repr<

' 1/99.

3.7 Anion Gap (AGP)


This derived test is calculated from Na + , CI" and CO 2 test results as follows: AGP = To access AGP processing: Press SHIFT, then SERVICE/CAL MODE to enter the service mode. You may only enter this option if there are no samples processing. If samples are processing, the analyzer will beep and display - WAIT - TESTS IN PROGRESS until it completes all tests. When the display prompts you to ENTER OPTION NO., key in 27 and press ENTER. Press 1, then ENTER to turn on AGP derived test processing. Press SHIFT, then SERVICE/CAL MODE again to exit the service mode. Now you are ready to run the component tests as outlined in section 2 of this manual, Operating Instructions. Once the patient ID is entered and Na + , CI" and CO 2 slides are analyzed, the AGP result will automatically be calculated and printed along with the Na + , CI" and CO 2 test results. If more than one value for Na + , CI" or CO 2 for a given patient ID appears in the last 20 results buffer, the test name and MULTIPLE RESULTS will appear on the printout followed by the name of the derived test and NO RESULT-COMPUTE MANUALLY. Na+-(Cr

5/95. Reprinted 1/99.

Operator's Manual . VITROS DT II System

3-33

This page has been left blank intentionally.

3-34

Operator's Manual VITROS M System

5/95. Repr

'1/99.

3.7 Anion Gap (AGP) (continued)


To turn anion gap processing off: Press SHIFT, then SERVICE/CAL MODE. When the display prompts you to ENTER OPTION NO., key in 27 and press ENTER. Press 0, then ENTER to turn off the AGP derived test processing. Press SHIFT, then SERVICE/CAL MODE again to exit the service mode.

1. Results from component test, Cl", CO 2 ,

and Na , followed by AGP result.

2. A 1-10 character patient ID was not entered prior to running Cl", CO 2 , and Na + tests.

3. The analyzer cannot calculate results for AGP if there are multiple results for any of the component tests.
* f * -i- . i- ,: * * * * * * * * * * .* *

01-29-92 ******************* CLSl MMOL-'L

*******************
I.D: 61-20-92 ******************* UL-

I.D:
* :f * f -:

2
* * * * * * * * * * * *

CL12: .iMOL-'L

32 MMOL/L C02 25 riMOL/L HA+ 119 MMOL/L


*******************
121

C02 22
NA+

COX

:M0L/L

119 MMOL/L ******************* I.D: NA+ BLANK PATIENT ID AGP NO RESULT COMPUTE MANUALLV *******************

NA +

I.D: NA+ CLC02


A8P

484243 119 MMOL-'L 81 MMOL.-'L 25 PHIOL-'L


13 MMOL-'L

***y- i .

. * * * * * * * * * * * *

I.D: 2 CLMl'-'-PLE RESULTS


AGP ..: RESULT
COM? W T M H N U A L L V
* * * . . * . , : * * * * * * * * * * * *

H e * * * * * * * * * * * * * * * * * *

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

3-35

This page has been left blank intentionally.

3-36

Operator's Manual .VITROS 'I System

5/95. Repr

'1/99.

Calibration
Calibration of blood-chemistry analysis equipment is an extremely important part of good laboratory procedure, since calibration helps to ensure the accuracy of test results. Faulty calibration or failure to calibrate the VITROS DT60 II System when recommended can lead to erroneous test results. When you calibrate the DT60 II System, you are programming new information into the software that translates data gathered during a slide reading into test results. Thus, during calibration, you are establishing new parameters for this translation process. The actual calibration procedure involves the analysis of VITROS calibrators fluids with known analyte concentrations in much the same manner as you analyze patient samples. Unlike sample analysis, however, the entire calibration procedure takes place with the System in the calibration mode, rather than the run mode. This procedure has been designed with ease and flexibility in mind. For instance, while it is suggested that you calibrate the tests in a certain order to make sure you do not miss any of them, you are free to calibrate in any order you wish. Thus, you can rearrange this procedure to meet your special needs.

4.1 Why You Need to Calibrate


Periodic calibration of the DT60 II System is required to maintain instrument reliability. As you calibrate, the analyzer establishes calibration parameters used to translate the response of the analyzer into concentrations of a desired analyte. These parameters, which are printed out at the completion of the calibration process, are stored in the analyzer's microcomputer memory.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

4-1

This page has been left blank intentionally.

4-2

Operator's Manual VITROS 'I System

5/95. Repr

'1/99.

4.2 When to Calibrate


Calibration is crucial to the ongoing reliability of your test results. CALIBRATE THE ANALYZER FOR ALL TESTS: 1. When the analyzer is initially installed. 2. When government regulations require.

- For example, in the USA, CLIA regulations require calibration or calibration verification at least once every six months.
3. When your Field Engineer indicates that calibration is necessary because servicing procedures might have affected the validity of the stored calibration parameters. Calibrate the analyzer for individual tests: 1. When the lot number of the ViTROS DT Slides change. 2. When the results of a quality control test using VITROS DT Controls or VITROS DT Isoenzyme Controls are consistently outside an acceptable range. 3. When a new lot of VITROS DT Reference Fluid is used. (This requires recalibration of tests run on the DTE II Module only.) NOTE: Refer to Section 13, "Log Sheets," for a sample of calibration log sheets to record data.

Rev. 2004-03-30

Operator's Manual VITROS DT II System

4-3

This page has been left blank intentionally.

4-4

Operator's Manual VITROS 'I System

5/95. Repr

M/99.

4.3 How to Calibrate


The major steps in the calibration procedure are: preparing the calibrators, analyzing them on the instrument, and assaying quality control fluids. Refer to the chart provided in this section for procedural steps on calibration. The procedure for analyzing the calibrators is designed to be simple and straightforward. In fact, it is very similar to the procedure used to analyze patient samples. However, the analyzer operates in the calibration (CAL) mode and analyzes the concentration of VITROS DT calibrators rather than patient samples.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

4-5

This page has been left blank intentionally.

4-6

Operator's Manual . VITROS 'I System

5/95. Repr

'1/99.

4.3.1 Preparing VITROS DT Calibrators


Calibrators needed for the DT60 II System are generally lyophilized materials that need to be reconstituted and used as soon as possible. Refer to the instruction sheet packaged with the product for reconstitution procedures and storage and stability requirements. STEP 1. Allow calibrators to warm to room temperature. 2. Open bottles. ACTION TO TAKE Allow at least 30 minutes for frozen calibrators and diluents. Remove metal seal from each bottle just before you intend to use them. IMPORTANT: DO NOT allow the diluents to stand without the stoppers. 3. Reconstitute the calibrators. Tap the top of each bottle so that any lyophilized materials adhering to the inside of the rubber stopper drop down into the bottle. Add exactly 3.0 mL of the appropriate diluent to each vial of lyophilized calibrator. Use a new 3.0 mL pipette tip for each diluent. Discard any remaining diluent. IMPORTANT: DO NOT interchange calibrators and diluents.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

4-7

This page has been left blank intentionally.

4-8

Operator's Manual VITROS 'I System

5/95. Repr'

i 1/99.

4.3.1 Preparing VITROS DT Calibrators (continued)


STEP ACTION TO TAKE

4. Dissolve all lyophilized particles.

Allow calibrators to stand for 30 minutes. Slowly invert the diluent bottles several times to mix the contents thoroughly. Swirl bottles periodically, but DO NOT SHAKE the bottles. Invert the bottles slowly to dissolve the lyophilized particles. All particles must be dissolved before using.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

4-9

This page has been left blank intentionally.

4-10

Operator's Manual VITRO!" II System

5/95. Rep

i 1/99.

4.3.2 Entering the Calibration Mode


STEP 1. Allow the slide to warm to room temperature. 2. Enter the calibration mode. 3. Enter the calibrator kit number. ACTION TO TAKE This takes approximately 1 5 minutes. Press the CAL key. The word CAL will appear in the upper right hand corner of the display. The display will prompt you ENTER CALIBRATOR KIT NUMBER. Enter the kit number that is printed on the carton containing the calibrators, then press the ENTER key. 4. Enter the generation number of the Reference Fluid. Enter the generation number found on the label of Reference Fluid, then press the ENTER key. If you do not have reference fluid, then enter 01 and press the ENTER key.

Rev. 2004-03-30

Operator's Manual VITROS DT II System

4-11

This page has been left blank intentionally.

4-12

Operator's Manual VITROS 'I System

5/95. Repr

M/99.

4.4 Calibration
STEPS FOR CALIBRATION O N THE VITROS DT II SYSTEM STEP GENERAL INFORMATION

1. Push slide into spotting station.

Run bottles in sequence. The display will prompt you which bottle numbers to use. Enter calibrator bottle number by pressing the bottle number and ENTER key. IMPORTANT: DO NOT insert slide until DT60 indicates "ANALYZER READY". IMPORTANT: DO NOT remove slide from spotting station after insertion unless directed by the analyzer.

2. Insert and fill dual-sample cup.

DTE II Module only. See Section 2 for specific instructions.

3. Aspirate calibrator fluid.

For specific instructions on proper pipetting techniques see Section 2. Carefully place pipette into calibrator bottle. DT60 II and DTSC II Aspirate fluid. Check fluid volume as shown in illustrations.

Fluid Level Check for Air Space at Tip

Rev. 2004-03-30

Operator's Manual VITROS DT II System

4-13

This page has been left blank intentionally.

4-14

Operator's Manual VITROS 'I System

5/95. Repr

< 1/99.

4.4 Calibration (continued)


STEPS FOR CALIBRATION O N THE VITROS DT II SYSTEM STEP 4. Spot slide. GENERAL INFORMATION

Carefully position pipette in locator. DT60 I I System With the pipette in the locator, depress and release the start button. An audible tone indicates that slide is spotted and you should then remove the pipette from locator. DTSC II Module A green flashing light and an audible tone indicate a "ready-to-spot" status. When spotting is completed the green light will go off. DTE II Module Depress the pipette button and continue to hold it depressed. With the button still depressed, slowly remove the pipette from the locator. An audible tone will tell you when the slide has been spotted. A flashing red light appears to indicate incubation has begun.

IMPORTANT: DO NOT remove slide from spotting station after insertion unless directed by the analyzer.

5. Eject tips.

DTE II Module Discard dual-sample cup after each bottle.

Rev. 2004-03-30

Operator's Manual VITROS DT II System

4-15

This page has been left blank intentionally.

4-16

Operator's Manual .VITROS' '[System

5/95. Repr

'1/99.

4.4 Calibration (continued)


STEPS FOR CALIBRATION O N THE VITROS DT II SYSTEM STEP 6. Repeat steps 1-5 for the same test using the next bottle. 7. Examine test results. GENERAL INFORMATION Flashing numbers on the display indicate the bottles which have been run for a particular test. Printout should read as follows: * 1 REP # 1 The number after the * is the bottle number, and the number after the # is the successful replicate completed. IMPORTANT: if a slide is not processed successfully on the first rep during calibration, a zero is reported instead of a valid replicate number (for example, * 1 REP # 0). If this happens, run another slide for the same test using the bottle number indicated on the display screen.

8. Repeat steps 1 through 7 for other tests.

Before exiting the calibration mode, be sure there are no printout messages which report a zero (0) instead of a valid replicate number.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

4-17

This page has been left blank intentionally.

4-18

Operator's Manual VITROS 'I System

5/95. Repr

' 1/99.

4.4 Calibration (continued)


STEPS FOR CALIBRATION O N THE VITROS DT II SYSTEM STEP 9. Exit calibration mode. GENERAL INFORMATION Press CAL key. If you accidentally forgot to run a bottle (for example, * 1 REP # 0), the display will read:

TEST MISSING BOTTLES LOAD "TEST" OR PRESS CAL TO EXIT.


Rerun missing bottle if indicated. If you do not run the bottle indicated, the test will not be calibrated. Press CAL key a second time to exit cal mode. Date and save calibration printout, label with 12digit slide lot #.

***************
CALIBROTION GLU CP*1-5.6943 CP#2 106.9S CP#3 2.3413 *************** *************** CALIBRATION URIC CPK1-1.8474 CP#2 12.551 CP#3 1.581S *************** *************** CALIBRATION K+ CFB1 1.2825 CP#2 .016945 ***************

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

4-19

This page has been left blank intentionally.

4-20

Operator's Manual VITRO? II System

5/95. Repr

11/99.

4.4 Calibration (continued)


STEPS FOR CALIBRATION ON THE VITROS DT II SYSTEM STEP 10. Run a quality control test. GENERAL INFORMATION Refer to Section 6.

IMPORTANT TIPS
Warm slides and calibrator fluids to room temperature. Do not interchange calibrators and diluents. For tests run on the DTE II Module, it is recommended that you run each bottle twice. Examine printout results. Run a quality control test to verify calibration.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

4-21

This page has been left blank intentionally.

4-22

Operator's Manual VITRO? II System

5/95. Rep'

11/99.

Instrument Care and Cleaning


Your VITROS DT60 II Chemistry System is designed to keep routine care and cleaning to a minimum. Since all analyses take place within VITROS DT Slides, there are no liquid reagents to mix or dispose of and fewer opportunities for messy spills. The self-contained nature of the analysis method also minimizes the need to clean the internal parts of the equipment. In short, the analyzer is not a demanding piece of equipment, but it does require some simple care to keep it operating reliably. The dry chemistry technology employed by DT Slides and the design of the DT60 II System, DTE II Module, and the DTSC II Module combine to minimize the need for care and cleaning. However, attention to daily and weekly cleaning, along with good housekeeping practices (keeping the analyzer and surrounding work areas as clean as possible, wiping up spills on the analyzer surface as soon as they occur) are recommended to provide for the continued optimum performance of your equipment.

CAUTION: Routine cleaning does not necessitate opening the analyzer's main cover.
Do not remove main cover or clean analyzer, DTE II Module or the DTSC II Module with ammonia or ammonia-containing compounds.

Rev. 10/01

Operator's Manual VITROS DT II System

5-1

This page has been left blank intentionally.

Rev. 1^""T

Operator's Manual VITROS r M System

5-2

5.1 Daily Cleaning


WHAT TO CLEAN 1. Slide disposal box(es). HOW TO CLEAN Lift out box(es) from analyzer and modules. Wash box(es) in a dilute solution of sodium hypochlorite for example, a 10% solution of liquid household bleach, or autoclave the box(es). Replace box(es) after cleaning. NOTE: Please refer to the VITROS DT Pipette User's Guide for information on cleaning and charging the DT Pipette.

Rev. 10/01

Operator's Manual VITROS DT II System

5-3

This page has been left blank intentionally.

Rev. 1 r "M

Operator's Manual VITROS ^ " M System

5-4

5.2 Weekly Cleaning


Clean the analyzer and modules as outlined here. You may use these same procedures when you detect accumulated dust or serum contaminants on the equipment during normal operation. Cleaning the DT60 II System WHAT TO CLEAN HOW TO CLEAN Clean the pipette locator and all visible slide track areas with a cotton swab moistened with water.

1. Pipette locator and visible slide track area.

Pipette Locator

Slide Track

Rev. 10/01

Operator's Manual VITROS DT II System

5-5

This page has been left blank intentionally.

Rev. T"

Operator's Manual VITROS' 'I System

5-6

Cleaning the DT60 II System (continued) WHAT TO CLEAN 2. Bar code reader and drop detector surfaces. H O W TO CLEAN Put your thumb in the hole at the front of the analyzer spotting station cover and lift up to expose the internal assemblies. Use lukewarm water and a clean, dry cotton swab to clean the surfaces. When cleaning is complete close the spotting station cover.

Bar Code Reader

Drop Detector Sensor

Rev. 10/01

Operator's Manual VITROS DT II System

5-7

This page has been left blank intentionally.

Rev.

Operator's Manual VITRO? 'I System

5-8

Cleaning the DTE II Module WHAT TO CLEAN 1. Pipette locator and visible slide track area. HOW TO CLEAN

Clean the pipette locators at the aspiration and spotting stations and the visible slide track areas, with a clean, cotton swab moistened with water. After cleaning, dry the area with another swab.
Pipette Locator

Slide Track

Pipette Locator

Rev. 10/01

Operator's Manual VITROS DT II System

5-9

This page has been left blank intentionally.

Rev. 1

'

Operator's Manual . VITROS 'I System

5-10

Cleaning the DTE II Module (continued) WHAT TO CLEAN 2. Rubber boot on the front of the electrometer. HOW TO CLEAN

Remove the pipette locator by placing your hand on the module and your thumb under the pipette locator slot. Lift the pipette holder up and out. Remove the nose assembly by turning the screw counterclockwise. Inspect and clean nose assembly and rubber boot with distilled water, then dry with lintless tissue. Place back into position and tighten screws clockwise.
1

Replace the pipette locator by placing the locator at an angle and placing the tabs on the square holes. Pull up slightly and push in and downward at the same time.
Electrometer Nose Assembly, Removed Top View

Screw

Bottom View Electrometer Assembly '

Rubber Boot

Rev. 10/01

Operator's Manual VITROS DT II System

5-11

This page has been left blank intentionally.

Rev. 1

Operator's Manual VITROS 1 System

5-12

Cleaning the DTSC II Module WHAT TO CLEAN 1. Pickup and slide spotting stations. HOW TO CLEAN Clean these stations with a clean, absorbent cloth moistened with water. Dry them with a cloth after cleaning. Raise the access cover of the module to expose the internal parts. Remove the pipette locator. Use water and a clean absorbent cloth to clean the length of the slide track. Dry the slide track after cleaning.

2. Slide track.

Slide Spotting Station

Pickup Station

Rev. 10/01

Operator's Manual VITROS DT II System

5-13

This page has been left blank intentionally.

Rev. 1

'

Operator's Manual VITROS '! System

5-14

Cleaning the DTSC II Module (continued) WHAT TO CLEAN HOW TO CLEAN Use water and a swab to clean the hard-to-reach areas. Carefully replace the pipette locator.

3. Pipette locator.

Disassembled Pipette Locator

Rev. 10/01

Operator's Manual VITROS DT II System

5-15

This page has been left blank intentionally.

;. Rev. '
1

Operator's Manual VITROS II System

5-16

Cleaning the DTSC II Module (continued) WHAT TO CLEAN HOW TO CLEAN The white reference cap is located on the underside of the read station preheater arm. Use warm water and a clean, absorbent cloth. The sapphire read window is directly beneath the read station arm. Use warm water and a clean, absorbent cloth to clean the window.

4. White reference cap and sapphire read window.

White
Reference Cap

Sapphire Read Window

Rev. 10/01

Operator's Manual VITROS DT II System

5-17

This page has been left blank intentionally.

Rev. 1

Operator's Manual VITROS 'I System

5-18

5.3 Other Cleaning: VITROS DT60 II Chemistry System FORS Head


Cleaning of the fiber optics reflectance system (FORS) head in the analyzer is recommended prior to recalibration and when dust is present. The analyzer will prompt you with one of the error codes R11-R1 7 to notify you of the necessity to clean. WHAT TO CLEAN FORS HEAD HOW TO CLEAN Put your thumb in the hole at the front of the analyzer's spotting station cover, and lift it to expose the internal assemblies. To expose the FORS head, lift up on the weight that covers the head and rotate it out of the way. Clean with a cotton swab moistened with water. Dry with a fresh cotton swab. Remove all lint. Return the weight into position over the FORS head and close the spotting station cover.

Weight Covering FORS Head

FORS Head

Rev. 10/01

Operator's Manual VITROS DT II System

5-19

This page has been left blank intentionally.

Rev. '

'

Operator's Manual . VITROS 'I System

5-20

5.4 Paper Loading 5.4.1 Removing the Paper


STEP 1. Remove printer cover. ACTION TO TAKE Position your thumb on the top of the cover and your fingers along the bottom edge of the cover where it joins the left side of the analyzer. Pull it up firmly and away from the analyzer. Slide the paper core out of the printer cradle and cut the paper tape at any point between the core and the slot where the paper enters the printer. After the tape is cut away from the core, remove the remaining tape by pulling it out through the print head. CAUTION: Remove the paper with a forward motion only. Reverse motion can damage the feeder.

2. Remove paper core.

Rev. 10/01

Operator's Manual VITROS DT II System

5-21

This page has been left blank intentionally.

Rev. 1

'

Operator's Manual VITROS II System

5-22

5.4.2 Inserting Paper


STEP 1. Load new roll. ACTION TO TAKE Cut both corners of the edge. Position paper roll as shown in illustration. Load paper into the printer cradle keeping the paper in the same position so that it feeds from the bottom of the roll.

Rev. 10/01

Operator's Manual VITROS DT il System

5-23

This page has been left blank intentionally.

Rev. 1 ^ 1

Operator's Manual VITROS 'II System

5-24

5.4.2 Inserting Paper (continued)


STEP 2. Feed paper into print head. ACTION TO TAKE Feed paper over the roller and through the slot in the back of the print head. Feed the paper until you feel resistance, a sign that the paper has engaged in the pressure roller of the print head.

3. Advance paper.

Press the PRINT key to feed the paper through the print head. If the paper does not feed through the print head, try feeding it through the slot in the back of the print head again, or it may be necessary to turn the DT60 II System off and back on again to reset the printer.

Rev. 10/01

Operator's Manual VITROS DT II System

5-25

This page has been left blank intentionally.

Rev. 1

'

Operator's Manual VITROS II System

5-26

5.4.2 Inserting Paper (continued)


STEP 4. Replace printer cover. ACTION TO TAKE Make sure that the paper feeds out through the slot between the printer cover and the rest of the analyzer surface. Manually feed the paper through the tear bar and paper cover.

Rev. 10/01

Operator's Manual VITROS DT II System

5-27

This page has been left blank intentionally.

Rev. 1

Operator's Manual VITROS 'I System

5-28

5.4.2 Inserting Paper (continued)


STEP 5. Check printer operation. ACTION TO TAKE Press the SHIFT key and then the SERVICE MODE key to enter the service mode. When the display prompts you to enter an option, enter Option 4. When the character set has printed out, press the SHIFT and SERVICE MODE key again to exit the service mode. The analyzer then returns to run mode.

Rev. 1 0/01

Operator's Manual VITROS DT II System

5-29

This page has been left blank intentionally.

Rev. 1 ^ 1

Operator's Manual VITROS^' II System

5-30

Quality Control
Reporting reliable patient test results can be assured through a comprehensive quality control program. Like calibration, regular quality control testing is an important part of laboratory procedures, playing a vital role in maintaining the accuracy of test results.

6.1 What Is Quality Control?


Quality control is a means of monitoring the precision and accuracy of the performance of an analytical system. In a clinical laboratory, an analytical system consists of: operators' techniques, the instrument, the reagents (for example, the slides), the calibrators and the environment of the laboratory, (for example, temperature and humidity). Precision is the reproducibility of a test and accuracy is a description of how closely your test results agree with the true value of the analyte being tested. Quality control verifies the success of a calibration and allows you to monitor the long-term performance of the equipment. There are two primary elements in maintaining quality control (QC). Monitoring of the control results Following generally accepted lab procedures

6.2 What Are Controls?


Controls contain known amounts of the same analytes that the analyzer measures in actual patient samples. This allows you to compare the results the analyzer delivers with known values. If controls fall within an acceptable range, patient samples may be analyzed with confidence.

6.3 How Often Should Controls Be Run?


Local regulations may indicate both the number of and frequency with which you are required to run controls. Analyzing controls daily will assist you in identifying changes in the system and taking action promptly. We recommend that, at a minimum, you perform a quality control test: Once a day for all tests being performed in your office that day After completing calibration When you suspect patient results are inaccurate After any major repairs performed on the analyzer In addition to these, your lab may establish its own requirements. It is important that the time interval between control tests satisfies both your needs and the requirements of local regulations.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

6-1

6.4 How to perform a quality control test 6.4.1 Preparing Lyophilized Controls
Most tests performed on the VITROS DT II System are lyophilized, (freeze-dried) material, although some are in liquid form. These are prepared from human and bovine serum and should be handled with the same precautions that you would with any other serum sample. The lyophilized controls must be reconstituted before use. Avoid ingesting any material and wipe up any spills immediately. Refer to the instruction sheet packaged with the product for storage and stability requirements. 1. Allow the controls to reach room temperature. Materials should be at room temperature before reconstitution. Vials should sit out approximately 30 minutes if stored in refrigerator or approximately 60 minutes if stored in the freezer. 2. Open the bottle. Remove the metal seal from the bottle. IMPORTANT: DO NOT allow the control or diluent to stand open without the stopper. 3. Reconstitute the control. Tap the top of the bottle to dislodge any particles before opening. Invert the diluent bottles several times to mix the contents thoroughly, but DO NOT SHAKE. Use a new pipette tip for each vial. Add exactly 3 ml_ of the appropriate diluent to the vial of the lyophilized control. Discard any remaining diluent. 4. Dissolve all lyophilized particles. Allow controls to stand for 30 minutes with occasional inversion, or place on a rotator or rocker for 10 minutes. Invert the bottles gently, but do not shake. Check that no undissolved particles remain before using. When not in use, cover the vial tightly, label with the date, and refrigerate. To best realize the keeping stability of the control, transfer the amount needed for the day's use from the vial into another container, and return the vial to the refrigerator. Seal the working container to prevent evaporation.

6-2

Operator's Manual VITROS DT II System

5/95. Reprinted 1/99.

6.5 Analyzing the Controls


1. Run the controls. Use the same procedure that you use when analyzing a patient sample, substituting the control for the patient sample. See Section 2, "Operating Instructions". When the display prompts you to enter the patient I.D., substitute the lot number on the bottle of the control fluid.

Rev. 2004-03-30

Operator's Manual VITROS DT II System

6-3

6.6 How to Record the Results


It is important to record the results of your control tests in order to monitor trends and patterns over a period of time. Data can be tracked on either of two forms: on a quality control log or on a graph.

6.6.1 Recording Quality Control Results on a Log


If you are using VITROS DT Controls you may find the necessary information on the yellow assay sheets or you may enter the values established by your lab.
VITROS CheratSTfysProducts DT Control I Assay Sheet Lot Number (T1317) CAT 842 0317
Expiration Dale: Auquft3i, 1997 / VITROS DT Systama

1 i mrnmm V i l u * aa" "Qp|| |v1u. Bang*


Analytt GEN AMVL 62/63 129 140 1.1 0.9 0.6 BUWJREA CHOL V 53 54 18 15-21 152-178 148 174 0.6-!.? 74-96 mfl/dL 1 ma/dL mgrtL mg/dl mg/dL 104-154 115=165 ' o . e - T6 0.4-1.4 \/L 29 40 IS 15 \ 10

Rang* 104-154 115-165 10-27 7-24 2-19 S.4-7.5

Units IM. U/L j*mo1/i_


LJTIOL'1

Ftafaranea Fluid Ganarallon 01 Anaiyta Gn CO ? Value 24.2 80 2.9 119 Ranga 19.2-29.2 75-B5 2.6-3.2 Unlla mmoi/L rwr.oi/L mmol/L

TBIL

16B f>((,'67

mfl/dtt mutiny

cr
K* Na +

mmDlA.

CREA GLU

AN "\
o\ 4.7 'U . 1-5.3

114 124 mmoK.

mmol/1. 53-106SsPmofL Control Log Slwet

317
iot no. changes, use the new comrot aooay find acceplabie ranges and begin e new

ol Value

Acceptable?

Corrective Action/Comments

12/12 12/13 12/14 12/15

0.9 1.0 1.8 1.0

'Jes 'Jes
9{o

Repeat test-1.4-0.%

Jes

TEC TEC TEC TEC

1. Record the control lot number, test name and range on the log sheet. Confirm that the control lot number on the assay sheet is the same as on the bottle of control you are using. 2. Record the date and result on the log sheet. 3. Compare the control value with the assay range on the log sheet. 4. If the control value is within range, it is acceptable. If it is not acceptable, see the "Quality Control Checklist". Record your corrective actions as indicated in the example. 5. Write your initials in the space provided. 6. Start a new log sheet when the control lot number changes. Record the new assay values on the control sheet.

6-4

Operator's Manual VITROS DT II System

5/95. Reprinted 1/99.

6.6.2 Recording Quality Control Results on a Graph


Charting quality control information on a graph such as the LeveyJennings is a simple way to present data for interpretation because it provides a visual representation of control results. The chart is based on the mean and standard deviation values which are calculated from a minimum of 20 control results for a given laboratory test. The mean is simply another term for the calculated average and is drawn as the center on the control chart; standard deviation (SD) is a statistical method for measuring dispersion. In terms of quality control, SD is used to evaluate how far quality control results fall from the mean. The acceptable range for control results is usually 2SD or 3SD. Because results may fall above or below the mean, SD is expressed as both a positive and a negative value. A blank Levey-Jennings chart is included in Section 13 of this manual. We have provided two charts so that you may monitor your normal and abnormal control results on a single sheet. If you are using VITROS DT Controls, you may use the mean (value) and range published on the control assay sheets. Confirm that the control lot number on the assay sheet is the same as the lot number on the bottle of control you are using. Using this information, fill in the Levey-Jennings chart.

VITROS Chemistry Products DT Control I Assay Sheet LotNumbe<jri317 CAT 842 0317
Expiration D a l e : AugusT3Trre37~"">y Conventions Value Units Range 129 140 1.1 0.9 0.6 (""is""1 53 S4 (M61 104-154 115-165 0.6-1.6 0.4-1.4 0.1-1.1 U/l U.t mg/dl mg/dL mg/dL mg/dL mg/dL mg/dL SI \ Value^ Range 129 140 19 15 10 6.4 4.3 4.2 VITiflOS DT Systems D T E M o d u l e s Reference Fluid Generation 01 Analyte Gen Value Range Units CO, 24.2 80 2.9
+

Analyte GEN AMYL 62/63 61/64/65 TBIl. NBIL BUN/UREA CHOI 68 66/67

Units U/L U/L

iWiS4 115-SfE

19.2-29.2 mmol/L 75-85 mmol/L 2.6-3.2 mmol/L 114-124 mmol/L

10-27 \ m t o l / L 7-24 (tfhoM. 2-19 5.4-7.5 3.9-4.6 3.8-4.5 (imolO mmoi/L \ m mo I/L mmo!/f

cr
K* SNa

) C^
152- 78 148- 74

Ms

119

Obtain from the side panel of your box of slides.

Limit Observed Value


r

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

6-5

1. Write the test name, generation number, slide lot number, control and control lot number in the spaces indicated above the appropriate chart. 2. Fill in the mean (value), +2SD and -2SD values from the assay sheet. To determine the SDs from the published assay sheet: +2SD = the upper range value listed on the assay sheet. -2SD = The lower range value listed on the assay sheet. 3. Fill in the +1SD and -1SD values, which must be calculated accordingly. +1SD = mean value + (+2SD value) 2 -1 SD = mean value + (-2SD value) 2 4. You are now ready to record results. Be sure to record the date and control test with each entry.

6.7 Interpreting Control Results 6.7.1 Interpreting the Control Results from the Log
If you are using VITROS DT Controls compare the printout with the results listed on the assay sheet that is packaged with the controls. Or you can compare the printout with the values established by your lab. If results are within range, it is acceptable. If results are outside the range, repeat the procedure for that particular test and consult the checklist at the end of this chapter.

6.7.2 Interpreting the Control Results from the Graph


Generally, a test is considered "in control" if the results fall within the 2SD range on the chart. If the control result falls outside the 2SD limit, there may be a problem that requires attention. Results may also fall out of range clue to random chance. In either case, simply repeat the test. If the control result falls back in range, the test is considered in control and you may proceed to report patient results. Other conditions that the chart will reveal to you are trends and shifts. A trend in a control chart refers to a series of control values that continue to increase or decrease over a period of several consecutive days. It may be indicative of an unsatisfactory laboratory condition. When values on several consecutive days fall on one side of the mean but remain at a constant level, the trend is referred to as a shift. Regular use of the Levey-Jennings chart will not only assist you in documenting your quality control activities, but can provide insight into the overall performance of each test for preventive actions before control values go out of range.

6-6

Operator's Manual VITROS DT II System

5/95. Reprinted 1/99.

This control chart indicates a trend in the observed value from 2/7 to 2/14, respectively.

'SUihi____ Gen # J 0 _ Slide Lots 0153-0223-4789 Control Vitro?'PI'ControlJ Control Lot #

'11317 Level 1

Date
Upper = Limit Mean = Lower _ Limit
91

-----

Observed Value
Corn ictive Action:

2/14/96

Level1:

Cleanedpipette/repeatedtest
__J&ity_!D_ay_ _.

Reviewed: Date __2/2&/96

Signature

This control chart indicates a shift in the observed value from 3/11 to 3/14, respectively.

Test '3itJ{ Gen# S3 Slidel_ot# 0_l53-0223-41S9

control VitmsDIComroll_ controlLots

T1317_ Level 1

Date
Upper = Limit Mean = _ Lower = Limit
21

15

corrective Action: Reviewed- Date

3/15/96 3/28/96

Level1: '-Reconstituted a new vial of control Signature 'Betty 'Day

6.8 Establishing Your Own Control Ranges


The determination of the target value for your laboratory is based on statistical methods. Whenever you test a quality control fluid containing a known quantity of a certain analyte, your test results will vary slightly due to such factors as: environmental differences, how the test was performed, and the performance of the analyzer. These variations are considered normal. However, it is important in assuring test accuracy that your results fall within the established range of the control, preferably close to the target value.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

6-7

6.8.1 How to Calculate the Mean


A mean (average) can be calculated on a calculator. The following is an example of how to calculate a mean for the last 10 quality control tests you have run, for example, on glucose: (For statistical analysis you may want to collect more data points.) llOmg/dL 120mg/dL lOOmg/dL 107 mg/dL 112mg/dL 120 mg/dL 103 mg/dL 110 mg/dL 104 mg/dL 114 mg/dL 1. To calculate the mean, add these values. The total is 1100. 2. Divide this number by the number of values quality control tests you obtained, in this case 10. Thus, your mean for glucose is 110 mg/dL (1100 + 10 = 110).

Operator's Manual VITROS DT II System

5/95. Reprinted 1/99.

6.8.2 Variables to Consider in Establishing the Mean


The values used to calculate the mean will affect the result. Although many commercially available control fluids have been tested to provide a pre-determined mean, you should establish your own mean target for each test. To establish such a mean, you should run quality control tests daily for a number of days. In hospital laboratories, at least 20 days of quality control results are gathered. Running QC tests for 20 days before establishing a mean target value for each test may not be feasible for an in-office laboratory, therefore you should obtain a mean that includes at least five to ten days of QC results. The mean that you establish for your laboratory should take into account a number of important variables so that it simulates the situations that occur during normal patient testing as closely as possible. For instance, day-to-day variables are critical. The results used to calculate the mean must have been gathered over several days to account for day-to-day changes in the environment and in instrument upkeep. If more than one person operates the analyzer, you must also account for operator-to-operator variables in establishing your mean. In other words, all the operators who run patient tests must have a portion of the QC test that you use in deriving the mean. Your mean should also include vial-to-vial variables. The QC results used to calculate the mean must be run using more than one vial of control fluid.

6.8.3 Calculating the Standard Deviation


Standard deviation is a statistical method for measuring dispersion. In terms of quality control, standard deviation is used to measure how far quality control results differ from the average that you established. You should establish a standard deviation for each test when you have established your mean. This standard deviation measurement can make use of the same values used in deriving the mean.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

6-9

6.8.4 How to Calculate a Standard Deviation


1. Calculate your mean as shown in section 6.8.1. In this case the mean was 110. 2. Calculate how much each of your values deviates, or differs from the mean. 110 mg/dL = mean, so deviation = 0 120mg/dL = mean+10 100 mg/dL = mean -10 107 mg/dL = mean -3 112 mg/dL = mean +2 120 mg/dL = mean+10 103 mg/dL = mean -7 110 mg/dL = mean, so deviation = 0 104 mg/dL = mean -6 114 mg/dL = mean +4 3. Then you square each of these deviations, which means that you multiply it by itself. o 22 = 0 10 = 100 -10 2 = 100 -32 = 9 22 = 4 102 = 100 - 7 2 = 49 02 = 0 - 6 2 = 36 4 2 = 16 4. Add all of the squared values together. 0 100 100 9 4 100 49 0 36 16 414

6-10

Operator's Manual VITROS DT II System

5/95. Reprinted 1/99.

5. Next, divide this sum by the total number of values minus one, in this case, 1 0 - 1 . You would divide by nine in this example. (414-9 = 46). 6. Finally, you take a square root of this value. This step is designed to "undo" the extra step that you took earlier in which you squared all the values. You can easily find the square root by using a pocket calculator. This square root of 46, which is written 46 equals 6.78 (approx.). In this example, 6.78 - one standard deviation. Generally, a test value is considered to be acceptable from a quality control point of view, if it is within two standard deviations of the mean that you have established. In the example given, then any quality control result for glucose that was within 6.78 x 2 or 13.56 mg/dL, of the mean of 110 would be considered acceptable. This would include any values between 96.44 (110- 13.56 = 96.44) and 123.56 (110+ 13.56 = 123.56). NOTE: The standard deviation you obtain should be similar in magnitude to those published by the manufacturer. A result that is much higher than stated by the manufacturer's figure can signal a system problem.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

6-11

6.9 Quality Control Troubleshooting Chart


If a quality control value is inaccurate follow these guidelines to correct the problem.

Factors to Consider
Slides Stored properly. Warmed sufficiently. Lot calibrated. Control How old is the control? If > 7 days, use a new one. Check assay sheet for stability. Were reconsitution procedures properly followed? Pipette Tip(s) inserted properly. Tip fill sufficient. Pipette removed at correct time after spotting. Check battery was battery low indicator on? Reference Fluid DTE II Module only. Stored according to requirements; usable for only 30 days after opening. Fluid warmed sufficiently. Check expiration date on carton. How long at room temperature in dual-sample cup? New VITROS Reference Fluid lot # requires calibration of Na + , K*, Cl", and Step 1. Rerun the control.

For DTE II Moduleleave Stored according to requirements. button depressed after Warmedj V V CA 1 1 1 I v spotting until sufficiently. you remove from locator.
V-^

co2.
2. If QC fails again, call your Customer Support Center.

6-12

Operator's Manual VITROS DT II System

5/95. Reprinted 1/99.

6.10 Factors to Consider When Your Results Are Out of Range


You may find that your quality control results occasionally fall outside the acceptable range you have established, even after you repeat the QC test; or you may become suspicious of the results you obtain when you run patient samples. Either condition should alert you to the need to check the factors: Instrument maintenance Housekeeping procedures Errors in labeling and transcription Centrifugation Pipetting technique Pipette performance Storage and handling of patient samples Storage and handling of test fluids and supplies Calibration Storage and handling of patient samples - Keep all samples tightly capped. - Frozen samples should be properly mixed when thawed. - Spin down samples before you store them and draw serum/ plasma off. - Store samples in the refrigerator (refer to Instructions for Use). - Warm refrigerated samples to room temperature before analyzing them.

Rev. 2003-10-01

Operator's Manual VITROS DT II System

6-13

6-14

Operator's Manual VITROS DT II System

5/95. Reprinted 1/99.

Options
Keyboard options are primarily used to check the function of the equipment after it is initially installed. They are also used in troubleshooting the equipment. For example, you may run a certain option to respond to an error message on the display. Some options are also used to obtain data that are stored in th.e microcomputer's memory - for instance, the type of reporting units for a test.

7.1 How to Run Options


To access the options you must be in the service mode of operations. Follow the sequence outlined below to enter the service mode from the run mode, and then to return to the run mode when you have completed running the desired options. To enter the service mode: Press SHIFT then SERVICE/CAL MODE. When the display prompts you to ENTER OPTION NO., key in the desired option number and press ENTER. To exit the service mode, press SHIFT then SERVICE/CAL MODE. NOTE: You cannot enter the service mode when the DT60 II system is processing samples. Wait until all samples are processed before executing options.

7.2 Options for the VITROS DT60 II Chemistry System


OPTION NUMBER 0 1 TERMINATING OPTIONS SLIDE TO INCUBATOR DESCRIPTION Terminates Options 1, 2, 6, 7, 8, 14, and 31. Activates the mechanism (upper rack) that transports a slide from the spotting station to the incubator. Use Option 0 to terminate. Advances the lower slide transfer mechanism (lower rack) to the correct position for a white reference reading. Use Option 0 to terminate.

POSITION WHITE REF AT READ

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

7-1

OPTION NUMBER 3 ENTER # OF CYCLES (199)

DESCRIPTION

Cycles the slide transfer mechanism. This option is selfterminating. To specify number of cycles: Key in the desired number of cycles. Press ENTER. Checks the operation of the printer by printing out the entire set of characters from A to Z and 0 to 9, and all punctuation marks. This option is self-terminating. The two rows of 40 black rectangles light up. Lights the green LED in the FORS head. Run Options 0, 7, or 8 to terminate. Lights the red LED in the FORS head. Run Options 0, 6, or 8 to terminate.
Lights the yellow LED in the FORS head. Run Options 0, 6, dr 7 to terminate. Tests the read-write memory O N D14(RAM) in the analyzer. This option reinitializes the analyzer. Rerun all tests for all slides that were in the analyzer at the time of the loss. This option is self-terminating.

PRINTER TEST IN PROGRESS

5 6

(Display lights up) GREEN LED TEST

RED LED TEST

YELLOW LED TEST

RAM CHECK-INITIALIZE O N D14

7-2

Operator's Manual VITROS DT II System

5/95. Reprinted 1/99.

OPTION NUMBER 10 PROM MEMORY CHECK (IS VALID or ERROR IN PROM or D13)

DESCRIPTION Tests the read-only memory (PROM). If the display reads: ERROR IN P R O M - t h e keyboard will not accept any further information until the problem is resolved. PROM OK - the option will be self-terminating at its completion. D13-PROM CHECKSUM ERROR - call your Customer Support Center. Displays current reporting units. Run Option 63 to select U.S. Units or Option 64 to select Standard International Units (S.I.)- This option is selfterminating. Prints out the last 64 characters, entered. This option is selfterminating. Displays the current version of software and the CDM number installed in your system.

11

U.S. UNITS IN EFFECT or S.I. UNITS IN EFFECT

12

LAST 64 KEYSTROKES

13

VERSION XX.X CDM XXXX ALL RIGHTS RESERVED 1995 COPYRIGHT OCD A/D READS TO PRINTER

14

This test is usually performed only at the request of service personnel. Prints the analog-todigital reading with each test result in the run, calibration or service mode. Run option 0 to terminate.

Rev. 2003-10-01

Operator's Manual VITROS DT II System

7-3

OPTION NUMBER 16 LED#? (GR =6, RD =7, YL =8, OFF =9)

DESCRIPTION Prints analog-to-digital reading of the light coming through FORS. The display will prompt you to enter the followfng data: ENTER STEP CNT - select the number of steps to move the lower rack assembly (-255 to +255). ENTER # OF READS - select the number of analog-todigital readings. Your selections will appear on the display as they are made. Date should be entered before routine analyses begin. Use the numeric keys to enter any 8 digits. The dashes must be entered between the numbers or the date will not be recognized. Press the ENTER key after you have entered all numerals and dashes. Do not press ENTER after each individual entry. The date will then be printed out with every patient ID for your recording of records. See Section 2.2 for when you should enter the date. When enabled, shows the amount of time left for slides in the incubator. Energize the analyzer again to delete this option. Prints out an ammonia result with a creatinine result, if DSCREA is run. Turns the capability to process Coronary Risk Classification on and off. Allows you to verify the status as on or off. Touch 1 to turn on and 0 to turn off.

17

ENTER DATE NN-NN-NN

18

ENABLE COUNTDOWN

22

REPORT NH 3 WITH CREATININE CRC PROCESSING ON/OFF

23

7-4

Operator's Manual VITROS DT II System

5/95. Reprinted 1/99.

OPTION NUMBER 24 CRC DATA INPUT EDITING

DESCRIPTION This option automatically turns CRC on and allows you to input the data necessary to run Coronary Risk Classification and to run "what if..." tests. See Section 3, "Coronary Risk Classification" for specific operating instructions. Turns the capability to process Globulin and Albumin/Globulin ratio calculations on and off. Allows you to verify the status as on or off. Touch 1 to turn on and 0 to turn off. Option 25 will remain on even if the analyzer is powered off and then on again, until the option is manually turned off. Turns the capability to process VLDL, LDL and CHOL/HDLC ratio calculations on and off. Allows you to verify the status as on or off. Turns the capability to processAnion Gap (AGP) calculations on and off. Allows you to verify the status as on or off. Touch 1 to turn on and 0 to turn off. Option 27 will remain on even if the analyzer is powered off and then on again, until the option is manually turned off.

25

GLOB AND A/G RATIO ON/OFF

26

LIPIDCALC.-VLDL, LDL AND CHOL/HDLC RATIO ON/OFF

27

AGP ON/OFF

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

7-5

OPTION NUMBER 30 SAV UPDATE SELECT TEST AND INDEX

DESCRIPTION Allows you to change the SAVs and the post-prediction values for any test. For SAVs: When the display prompts you with SELECT CHEMISTRY, press the CHEMISTRY SELECT key to advance to the desired test. Press 1, 2, 3, or 4 to access the values for bottles 1, 2, 3, or 4. To change values, press the CLEAR key and enter the new value. To cancel, press SERVICE/CAL MODE. For post-prediction values: Press 6 to display the postprediction slope. Press 7 to display the postprediction intercept. To change the values, press the CLEAR key and enter the new value. To cancel, press SERVICE/CAL MODE. Use this option after using option 30 to change SAVs and before calibrating. Use Option 0 to terminate once calibration is complete. Allows you to enter and print calibration parameters. Press the CHEMISTRY SELECT key to scroll to the desired test. Press 1, 2, 3, and 4 for calibration parameters. Press 5 for the web generation number. To update the value, press the CLEAR key then enter the new value, and press ENTER. To print values, press the PRINT key. Press SERVICE/CAL MODE to terminate the option.

31

CDM DATA NOT USED: UNIT WILL USE EA PROM CAL VALUES

32

CALIBRATION UPDATE SELECT CHEMISTRY AND INDEX

7-6

Operator's Manual VITROS DT II System

5/95. Reprinted 1/99.

OPTION NUMBER 33 SEL-F1ELD, ENT-VALUE, PRINTSO WB BF (Access spline values off the CDM)

DESCRIPTION This option should be used only when recommended by trained OCD service personnel. This option is used in conjunction with Option 35 to print a selected group of spline records from the CDM. For the spline records, SO number, web generation, and body fluid code are used in the record system. Run Option 35 first. Press the CHEMISTRY SELECT key to move from one field to another. Select the field and use 0-9 for A-J, and SHIFT 0- SHIFT 9 for K-T. The CLEAR key clears any previous values. Press PRINT to obtain all t spline records. This option should be used only when recommended by trained service personnel. Run Option 35 first. Press the CHEMISTRY SELECT key to move from one field to another. Select the field and use 0-9 for A-J, and SHIFT 0-SHIFT 9 for K-T. The CLEAR key clears any previous values. Press PRINT to obtain all SAV values. The default for the Options 33 and 34 is the short form. Press 0 to obtain a long form. Prints the correction factors and reference slide reflectances. NOTE: Use Option 50 to obtain the data for the DTSC II Module.

34

SEL-FIELD, ENT-VALUE PRINTSO WB BF KN (Access SAV calibrator values off the CDM)

35

CDM LISTING O=FULL 1=SHORT PRINTING REFERENCE FACTORS (for DT60 II)

36

Rev. 2003-10-01

Operator's Manual VITROS DT II System

7-7

7.2.1 Serial Communication Options for the DT60 II System


OPTION NUMBER 19 SERIAL COMMUNICATIONS TEST DESCRIPTION This option is used to check the data transfer from the DT60 II System to an attached computer system. TEST OK - A record was successfully sent out and an acknowledgment was received by the external PC. UNABLE TO TRANSMIT The test record was not transmitted. NO RESPONSE FROM PC No acknowledgment was received from the PC. This option runs an interface test. Plug an interface connector onto the end of the serial communications cable. TEST OK - All characters sent and received correctly. TEST FAILS - The characters are not sent or received correctly. This option runs an internal loopback test. TEST OK - All characters are sent and received correctly. TEST FAILS - The characters are not sent or received correctly.

20

DATA INTERFACE LOOPBACK TEST

21

INTERNAL LOOPBACK TEST

7-

Operator's Manual VITROS DT II System

5/95. Reprinted 1/99.

7.3 Options for the VITROS DTE II Module


OPTION NUMBER 40 DTE ELECTROMETER FORWARD or ELECTROMETER AT READ DTE ELECTROMETER REVERSE or ELECTROMETER AT HOME ENTER # OF CYCLES DESCRIPTION

Advances the electrometer to the read position. Run Option 41 to terminate the option. Returns the electrometer to the home position. This option is self-terminating. Cycles the drive mechanism a specified number of times. Key in the desired number of cycles and press ENTER. This option is self-terminating Verifies the electrical performance of the electrometer. The gain factor, reference, and offset values of the electrometer will be the following range: GAIN FACTOR 24.0 to-21.5 REFERENCE 80.22 to-83.46 OFFSET 01 This option is self-terminating. Checks the incubator in the module. This test takes 40 seconds from the time you select the option. The following display messages may appear: DTE MODULE TEMP OK - temperature is within the correct range. DTE MODULE TEST FAILED - temperature is outside the proper range.

41

42

43

DTE ELECTOMETER VERIFIED

44

DTE MODULE TEMP TEST STARTED

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

7-9

7.4 Options for the VITROS DTSC II Module


OPTION NUMBER 50 DTSC REFERENCE VALUES (prints correction factors for DTSC II) DESCRIPTION The black and white correction values and reference slide values are printed out. Values are printed only for the filters that are in use. Each value will be labeled with its corresponding wavelength. Prints out the temperature in degrees Celsius. Prints out the current software version number.

52 53

DTSC TEMPERATURE DTSC SOFTWARE VERSION XX.X

7-10

Operator's Manual VITROS DT II System

5/95. Reprinted 1/99.

Troubleshooting
The software of the VITROS DT60 II Chemistry System, VITROS DTE II Module, and VITROS DTSC II Module monitors many hardware functions. Information regarding most sources of malfunction is provided to you through the display or the paper printout. This section provides you with instructions on what to do when your analyzer is not operating properly, including suggestions on the possible causes.

8.1 General Troubleshooting


Each description in this section lists one or more possible causes and a series of operator responses. Perform the steps in the order indicated. If one step does not correct the problem, proceed to the next one, and so forth, until all suggested responses have been completed. If the analyzer is not properly operating after all the suggested responses have been completed, contact your Customer Support Center. POSSIBLE CAUSE VITROS DT60 II System cannot be turned on. Power cord pulled from the analyzer or from the outlet. Main power switch in the OFF position. Nonfunctioning outlet. OPERATOR RESPONSE Verify that the power cord is plugged securely into the power outlet and the analyzer. Verify that the main power switch at the back of the analyzer is in the ON position. Verify that the outlet is functioning. If the analyzer is still not functioning, call your Customer Support Center. Check that the cable leading from the analyzer to the DTE II Module is securely plugged in. Turn the analyzer off, then on again. If the problem recurs, call your Customer Support Center.

VITROS DTE II Module is not operating.

Cable leading from module to the analyzer is not securely fastened. Instrument malfunction.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

8-1

POSSIBLE CAUSE VITROS DTSC II Module is not operating. Cable leading from module to the analyzer is not securely fastened.

OPERATOR RESPONSE Check that the cable leading from the analyzer to the DTSC II Module is securely plugged in. Turn the analyzer off, then on again. If the problem recurs, call your Customer Support Center. Check that the plug is plugged securely into the outlet. Turn the switch to the ON position. Verify that the outlet is functioning. See Section 5.6.2, "Paper Loading," for correct loading procedures.

Power cord is not securely plugged into the outlet. Main power switch is. OFF. Nonfunctioning outlet. Printer working but does not print hard copy. Printer paper inserted with heat sensitive side on the back. The heat sensitive side of the paper marks when scratched with finger nail. Main power switch in OFF position. Power cord pulled from the power outlet.

Complete power loss.

If the main power switch is in the OFF position, turn it to the ON position. Verify that the power cord is plugged securely into the analyzer and into the power outlet. Turn the main power switch to the OFF then ON again. Rerun tests for all slides that were in the analyzer at the time of the power loss. Repeat the calibration process for all tests being calibrated at the time of the power loss. If the problem recurs, call your Customer Support Center.

3-2

Operator's Manual VITROS DT II System

5/95. Reprinted 1/99.

POSSIBLE CAUSE Temporary power loss. Line voltage dropped below specifications.

OPERATOR RESPONSE If the voltage temporarily drops and then returns to normal, the unit will reinitialize. After reinitialization, repeat tests for all the slides in the analyzer at the time of the power loss. Repeat the calibration process for all tests being calibrated at the time of the power loss.

8.2 Pipette Troubleshooting


Please refer to the VITROS DT Pipette User's Guide for information on cleaning and charging the DT Pipette.

8.3 Unexpected Results (Analyzer Tracking Errors)


Tracking errors occur when the sequence of slides in the DT60 II System does not match the sequence the analyzer was expecting. Tracking errors are caused when display prompts are not followed exactly or when the slide disposal box is overfilled. Tracking errors cause the analyzer to report unexpected results such as: A series of results outside the analyzer range when running multiple chemistries on the DT60 II System. A series of highly unusual results outside the expected physiological range. Occasional slide transport malfunctions that are reported as F12 F15 error codes - "Transfer malfunction." Calibration failures that are reported as C13 (EXTREMA) or C18 (MONOTONICITY) error codes. Possible Causes A slide is pushed into the analyzer prematurely when the display shows a WAIT prompt. This may cause a transfer malfunction. A slide is not removed from the analyzer when the display prompts to do so. An example is an unreadable bar code on a slide that causes the analyzer to reject the slide.

Rev. 2004-03-30

Operator's Manual VITROS DT II System

8-3

An incorrect usage of the delete test key. The delete test key deletes a test after a slide has been entered, identified, and spotted. When a test is deleted, the slide must continue through the analyzer. Do not remove a slide manually once it has been deleted. IMPORTANT: If a slide is removed manually after the delete test is requested, a tracking error will occur. A slide disposal box that is not emptied on a daily basis. An overfilled slide disposal box causes the slides to backup resulting in a tracking error. If You Suspect a Tracking Error 1. Wait until the analyzer has finished processing. 2. Empty the slide disposal box. 3. Enter the service mode by pressing SHIFT then SERVICE/CAL MODE. 4. When the display prompts you to ENTER OPTION NO., press 3 then press ENTER. 5. When the display prompts you to ENTER # OF CYCLES (1-99), press 6 then press ENTER. 6. After the cycles have completed, check the slide disposal box. IMPORTANT: If the slide disposal box does not contain exactly one slide then a tracking error has occurred. 7. Exit the service mode by pressing SHIFT then SERVICE/CAL MODE. 8. The analyzer is now ready to resume routine operation.The above actions have cleared the tracking error. Please review and confirm previous results. Important Points to Remember Please follow the display messages. Insert and remove slides only when the display indicates to do so. Once the slide is spotted, use the SHIFT + DELETE keys to inhibit results from printing, but do not remove the slide.

8-4

Operator's Manual VITROS DT II System

Rev. 2004-03-30

Instrument Status Messages


Status messages may appear on the display of the printout during the course of normal operation for instance, in response to information that you input or in response to actions such as inserting a slide. This section includes a list of all messages along with explanations of the messages and recommended responses to them.

9.1 Status Messages


The messages in this section, for the most part, do not point to equipment malfunction. Instead they are designed to "talk you through" various operating procedures. MESSAGE ANALYZER INITIALIZATION (Display) ANALYZER READY-CREA/NH3 ONLY (Display) EXPLANATION The DT II System is going through a series of selfchecks. An N H 3 slide is in the incubator. ACTION TO TAKE Wait until the message disappears before attempting to run any tests. You must run a creatinine or ammonia slide next. To run any other tests wait until the creatinine and ammonia slides clear the incubator. Run a slide other than a creatinine or ammonia slide.

ANALYZER READY NO CREA/NH 3

In the CAL mode, creatinine and ammonia cannot be run while a slide for BUN is in the incubator. Instrument is still warming up.

ANALYZER TEMP. LOW (Display)

If the message appears when the analyzer is first turned on, wait until ANALYZER READY message is displayed (approximately 25 minutes). Check to see that the pipette locator cover is closed properly. After it is closed, the INCUBATOR WARMING UP message will appear. Wait until the display disappears and the ANALYZER READY display appears before running the next test.

COVER OPEN (Display) COVER OPEN TEST INVALIDATED (Printout)

Pipette locator cover on the analyzer was left open longer than 5 seconds.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

9-1

MESSAGE DISCARD PM SLIDE AND REPEAT (Display)

EXPLANATION Bar code reader cannot read the bar code on slide. Slide inserted upside down. The patient sample and electrolyte reference fluid were dispensed without first loading a slide.

ACTION TO TAKE Discard slide and repeat the test. This message will be displayed until the cover is lifted. Clean the barcode reader. Remove pipette locator cover. Examine internal assemblies and clean any serum or electrolyte reference fluid that may be present, following the instructions in Section 5.3. Wait until you hear the electrometer stop cycling, then run the desired option. Using the numeric keys on the keyboard, enter the number of the calibrator kit that you are using. This number is found on the box of calibrator fluids. Using the numeric keys on the keyboard, enter the level (1, 2, 3, or 4) of calibrator that you are about to use. Enter the generation number. If you do not have the DTE II Module, enter the number 1.

DTE ELECTROMETER CYCLING (Display) ENTER CALIBRATOR KIT NUMBER (Display)

A DTE II Module option was requested, and the electrometer is still cycling. The analyzer is prompting you to enter the number of the calibrator kit. This message appears after you enter the calibration mode. The analyzer is prompting you to enter the bottle number of the calibrator. The analyzer is prompting you to enter the reference fluid generation number on the bottle of VITROS DT Reference Fluid. The analyzer is prompting you to enter the generation number of the slide. This message appears after a test is selected in response to the SELECT TEST message.

ENTER CALIBRATOR BOTTLE (Display)

ENTER REF FLUID GEN NO. (Display)

ENTER SLIDE GENERATION (Display)

Using the numeric keys on the keyboard, enter the generation number of the slide that you just inserted. The generation number can be found on the box of slides being used. (The ENTER key must be pushed twice to identify the generation on the DTSC II Module).

9-2

Operator's Manual VITROS DT II System

5/95. Reprinted 1/99.

MESSAGE EXIT SERVICE MODE FIRST (Display)

EXPLANATION You tried to run a test while the instrument is in the service mode. This message is for service use only.

ACTION TO TAKE Exit the service mode by pressing the SHIFT and SERVICE keys on the keyboard. No operator response. Wait until message disappears or until the analyzer processes at least one test result before entering another slide. If you had already entered a slide, lift the pipette locator cover and remove the slide to prevent a jam. Close the cover within 5 seconds or the analyzer will reinitialize.

GAIN CHANGES INCUBATOR FULL (Display)

Incubator is filled to capacity.

INCUBATOR WARMING UP (Display)

The incubator in the analyzer has not yet warmed to the proper operating temperature. This message is displayed after spotting a creatinine slide to indicate that the next slide to be run must be an N H 3 slide. Printed as a result of a slide jam. The message that follows on the printout (F11-F15 TRANSFER MALFUNCTION) indicates the location of the jam.

Wait until the ANALYZER READY message appears before you attempt to run any tests. Load a slide for

INSERT NH3 SLIDE (Display)

OPEN COVER TO CLEAR (Printout)

Open the pipette locator cover and remove the jammed slide. When you close the cover, the instrument will reinitialize. Wait for the ANALYZER READY message to appear before running another test.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

9-3

MESSAGE REMOVE CM SLIDE (Display and Printout)

EXPLANATION You tried to spot a slide that was not identified. Bar code reader cannot read bar code on slide. You dispensed the patient sample without first loading a slide.

ACTION TO TAKE Check to be sure the slide is inserted into the correct module. Open pipette cover and remove slide within 5 seconds by pulling it back from the ends. (Do not touch center of slide because i t . contains the reagent). NOTE: If the pipette cover is open longer than 5 seconds, the analyzer will delete all slides in the incubator and require an incubator warm up. If a slide is being read at the same time that the cover is open, that slide result may be deleted. Reinsert the slide at a faster rate of speed. If the bar code reads, continue processing. If the bar code does not read, open the pipette locator and clean the bar code sensor with a dry swab. After the analyzer displays READY, reinsert the slide. If the bar code reads, continue processing slides. If the bar code still does not read, call your Customer Support Center.

9-4

Operator's Manual VITROS DT II System

5/95. Reprinted 1/99.

MESSAGE RESULTS ABOVE ANALYZER RANGE (Printout)

EXPLANATION In the calibration mode, this message informs you that the result from the calibrator fluid is above the range of the analyzer. Incorrect information was manually entered. In the run mode, the sample concentration is above the analyzer range.

ACTION TO TAKE Rerun the calibrator fluid in question. If message recurs, reconstitute a fresh calibrator fluid and rerun the test. If message recurs, call your Customer Support Center. If the CHEMISTRY SELECT key was used, make sure that all the information entered was correct. Dilute the sample and rerun the test. (Be sure to adjust the results according to the dilution factor). See the Instructions for Use for the test you are running for more information on dilution procedures. If message recurs, call your Customer Support Center.

RESULTS BELOW ANALYZER RANGE (Printout)

In the calibration mode, this message informs you that the result from the calibrator fluid is below the range of the analyzer. In the run mode, the sample concentration is below the analyzer range.

" Rerun the calibrator fluid in question. If message recurs, reconstitute a fresh calibrator fluid and rerun the test. If message recurs, call your Customer Support Center. If the CHEMISTRY SELECT key was used, make sure that all the information entered was correct. Rerun the test. If message recurs, run a quality control test. If quality control results are within the proper range, report the test results as being below the analyzer range.

Rev. 2003-10-01

Operator's Manual VITROS DT II System

9-5

MESSAGE SELECTING DTSC TEST (Display)

EXPLANATION This message is asking you to verify your test selection.

ACTION TO TAKE Press ENTER to verify the selection. If the selection is incorrect, use the CHEMISTRY SELECT key to scroll to the correct test and then press ENTER. Do not remove the unidentified slide. Leave it in the instrument and use the CHEMISTRY SELECT key to manually select the test that you have loaded into the spotting station. If the analyzer prompts you to remove the slide, do so as quickly as possible. If the cover is open more than 5 seconds, it could result in the loss of the tests in progress. If any of these problems recur, call your Customer Support Center.

SLIDE NOT IDENTIFIED (DT60 II System) (Display)

Slide was inserted backward. Instrument was unable to read the bar code. Unsteady motion while inserting the slide. Instrument malfunction.

9-6

Operator's Manual VITROS DT II System

5/95. Reprinted 1/99.

MESSAGE SLIDE NOT IDENTIFIED (DTE II Module) (Display)

EXPLANATION Slide was inserted backward. Instrument was unable to read the bar code. Instrument malfunction.

ACTION TO TAKE Visually check to see that the slide is correctly positioned below the pipette locator at the spotting station. The slide should be positioned as indicated in the diagram on the slide track. (The two small holes where the sample and reference fluid are deposited should face you.) If the slide is positioned correctly, press the CHEMISTRY SELECT key to manually identify the test. If the slide was inserted backward, insert a new slide. You can do this even while the SLIDE NOT IDENTIFIED message appears on the display. If the message remains, call your Customer Support Center.

TEST DELETED (Display and Printout) TESTS INVALIDATED (Printout)

The test was deleted by the operator before it was complete. The pipette locator cover was left open longer than 5 seconds, so the incubator temperature dropped and the tests in progress were invalidated.

Leave slide in and repeat test.

Check to see that the cover is closed. Wait for the ANALYZER READY message to appear, then repeat the test in question.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

9-7

MESSAGE TEST MISSING BOTTLES LOAD XXX OR PRESS CAL TO EXIT (DT60 II System) (Printout and Display)

EXPLANATION This message appears when exiting the calibration mode. The analyzer is indicating that you accidentally forgot to run a bottle during the calibration procedure, or you had an invalid calibration reading.

ACTION TO TAKE Load a slide for the test indicated to find out which calibrator bottle you are missing. Any number that is not flashing indicates the bottle that you need to run. After running the missing bottle(s) and after valid replicate numbers are printed, press the CAL key to exit the calibration mode. Wait until this message is replaced by the ANALYZER READY message before trying to insert the next slide.

WAITSLIDE BEING LOADED (Display)

The analyzer is indicating that the slide you just spotted with fluid on the DT60 II has not yet been transferred into the incubator.

IMPORTANT: DO NOT insert slide until the DT60 indicates "Analyzer Ready".
XXX (test name) USES BOTTLES (1,2,3,4) ENTER BOTTLE NO. (Printout and Display) During the calibration procedure, after the bar code on the slide is read, the analyzer is prompting you to choose the calibrator bottle that you want to run. To respond, enter the bottle number (press 1 , 2 , 3 , 4) to indicate which calibrator bottle you have chosen to run.

9-8

Operator's Manual VITROS DT II System

Rev. 2004-03-30

Coded Warning Messages


In addition to monitoring operating procedures, the analyzer's microcomputer monitors several analyzer functions and alerts you by means of coded warning messages on the display panel and/or the paper printout if there is a deviation from performance specifications. This section lists these coded messages in alphabetical and numerical order.

10.1 Coded Warning Messages


The code number beside each message appears on the printout and/ or the display along with the message. It is designed to help you locate troubleshooting information in this manual. The code number also provides valuable information to your Customer Support Center in case the instrument fails to operate properly after you have tried all the suggested responses listed here. If the problem persists, call your Customer Support Center.

10.1.1 Calibration (C)


CODE/MESSAGE C12 INVALID CALIBRATION (Printout) EXPLANATION/CAUSE This message appears as a heading on the printout. It is followed on the printout by another coded warning message that specifies the problem. Response (DR value) obtained for a calibrator bottle is not within an acceptable range: - bottles misidentified. - drop is too small. - calibrator bottle is reconstituted improperly. - Tracking error may have occurred ACTION TO TAKE Respond as instructed for the coded warning message that follows this heading.

C13

EXTREMA ERROR

(Printout)

Exit cal mode.To clear slides out of incubator, empty the slide disposal box and run option 3 (6 cycles) (VITROS DT60 II Chemistry System only). After the cycles have completed, check the slide disposal box. If the slide disposal box does not contain exactly one slide then a tracking error has occurred. Refer to section 8.3. Be sure pipette is clean. Repeat calibration with same calibrators. If the same error occurs, reconstitute new calibrators and recalibrate.

Rev. 2004-03-30

Operator's Manual VITROS DT II System

10-1

CODE/MESSAGE C14 SLIDE GENERATION ERROR (Display)

EXPLANATION/CAUSE Run Mode: Generation of slide loaded in the run mode has not yet been calibrated. Operator keyboard error (if number was entered manually). Calibration Mode: Incorrect calibrator kit for the test being calibrated.
o

ACTION TO TAKE Remove the slide and calibrate the test. Remove the slide and enter correct gen. #.

Use the correct calibrator kit: Calibrator to Use


VITROS Specialty Calibrator Kit VITROS Isoenzyme Calibrator Kit VITROS Calibrator Kit

Test
*Theo&CHE *CKMB All Other

*L19 error code will occur on DTSC II Module. Calibration Data Module (CDM) does not contain the information required for the slide generation being used. C15 KIT NUMBER NOT IN MEMORY (Display) Calibrator kit expired. Replace with the most current CDM. Install new CDMs when they arrive.

Check the expiration date on the calibrator box. Obtain new calibrator kit, if expired. Verify that the number entered was correct. Re-enter if necessary. Replace with most current CDM. Be sure to install CDMs when they arrive.

Calibrator kit not on CDM in the analyzer.

C17

MISSING BOTTLE (Display)

Calibration mode was exited before all levels were run for a given test.

Recalibrate the test in question. Follow proper procedure for all required levels of calibrator. If message recurs, call your Customer Support Center.

10-2

Operator's Manual VITROS DT II System

Rev. 2004-03-30

CODE/MESSAGE C18 MONOTONICITY ERROR (Print out)

EXPLANATION/CAUSE Response (DR value) obtained for a calibrator bottle is not within an acceptable range: - bottles run out of order. - drop is too small. - calibrator bottle is reconstituted improperly. - Tracking error may have occurred

ACTION TO TAKE Exit cal mode. To clear slides out of incubator, empty the slide disposal box and run option 3 (6 cycles) (VITROS DT60 II Chemistry System only). After the cycles have completed, check the slide disposal box. If the slide disposal box does not contain exactly one slide then a tracking error has occurred. Refer to section 8.3. Be sure pipette is clean. Repeat calibration with same calibrators. If the same error occurs, reconstitute new calibrators and recalibrate.

C19

CDM RECORD FORMAT ERROR (Display)

The computer software will not accept the record format number of the CDM. Damaged CDM or CDM socket. Software problem.

Turn analyzer off. Remove the CDM and reinsert it, or replace with a new CDM. Turn analyzer on and wait for it to reinitialize. If message recurs, call your Customer Support Center. Check the number on the reference fluid bottle label. Either replace the CDM or the reference fluid to achieve a match. Check that the number entered was correct and reenter if necessary. If message recurs, all your Customer Support Center.

C21

INVALID REF FLUID NO. (Display)

Calibration data module (CDM) is not programmed with the generation number of reference fluid that you entered. Operator keyboard error.

C22

NO N H 3 CALIBRATION (Printout)

The analyzer is trying to calibrate for creatinine, but cannot find the ammonia result, which should have already been calculated.

Recalibrate creatinine and ammonia as stated in the manual.

Rev. 2004-03-30

Operator's Manual VITROS DT II System

10-3

10.1.2 Data Storage (D)


CODE/MESSAGE D12 D13 EA PROM ERROR (Display and Printout) PROM CHECKSUM ERROR (Display) EXPLANATION/CAUSE Occurs when there is a computer malfunction. Occurs when there is a computer malfunction, either during initialization or while running option 10. Occurs when a memory check is unsuccessful. The correction factors are lost. Run option 36 (DT60 II System) or option 50 (DTSC II Module) to see if correction factors are in memory. If the correction factors are 0, enter the correction factors from a previous option 36/50 tape using option 81 (DT60 II System) or 101 (DTSC II Module). To determine if calibration data are in memory: - enter option 32. - press CHEM SELECT until desired chemistry appears on the display. - press PRINT. - compare the printout to the calibration data from a previously saved printout. Enter the lost calibration data again or wet calibrate the DT60 II System, DTSC II Module, or DTE II Module. Refer to option 32. Wet calibrate all chemistries if unable to restore lost calibration from saved printout.

ACTION TO TAKE
Call your Customer Support Center.

D14

RAM MALFUNCTION
(Display)

D16

MEMORY ERROR (Printout)

The calibration data are lost.

There was an error in calibration.

10-4

Operator's Manual VITROS DT II System

5/95. Reprinted 1/99.

CODE/MESSAGE D18 D19 MEMORY RESET (Printout) MEMORY HAS BEEN ERASED (Printout)

EXPLANATION/CAUSE

ACTION TO TAKE Call your Customer Support Center. Run option 36 (DT60 II System) or option 50 (DTSC II Module) to see if correction factors are in memory. If the correction factors are 0, enter the correction factors from a previous option 36/50 tape using option 81 (DT60 II System) or 101 (DTSC II Module). To determine if calibration data are in memory: - enter option 32. - press CHEM SELECT until desired chemistry appears on display. - press PRINT and compare the printout to the calibration data from a previously saved printout. Enter the lost calibration data again or wet calibrate the DT60 II System, DTSC II Module, or DTE II Module.

Instrument malfunction. The correction factors are lost.

The calibration data are lost.

There was an error in calibration. D25 DTSC INTERNAL RAM FAILURE (Printout) When initializing the DTSC II Module the software detects that the internal memory test has failed.

Wet calibrate all chemistries if unable to restore lost calibration. Turn the DTSC II Module off and then back on again, waiting for it to reinitialize. Repeat any tests that may have been deleted by turning the module off. If the problem recurs, call your Customer Support Center.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

10-5

CODE/MESSAGE D26 DTSC EXTERNAL RAM FAILURE (Printout)

EXPLANATION/CAUSE

ACTION TO TAKE

When initializing the DTSC II Module, the software detects that the external memory test has failed.

Turn the DTSC II Module off and then back on again. Wait for it to reinitialize. Repeat any tests that may have been terminated by turning the module off. If the problem recurs, call your Customer Support Center.

D27

DTSC BATTERY RAM FAILURE (Printout)

The DTSC II Module fails to write to the battery backed RAM as expected. This error can occur at any time during operation. When initializing the DTSC II Module, the software detects a checksum error on its PROM.

D28

DTSC CHECKSUM . FAILURE (Printout)

10.1.3 Electrometer (E)


CODE/MESSAGE E11 RESULTS INVALID (Printout) EXPLANATION/CAUSE ACTION TO TAKE

Instrument malfunction.

Check the interface cable. Repeat the test in question. If message recurs, callyour Customer Support Center.

E12

RESULTS INVALID (Printout)

Instrument malfunction.

10-6

Operator's Manual VITROS DT II System

5/95. Reprinted 1/99.

CODE/MESSAGE E13 RESULTS INVALID (Printout)

EXPLANATION/CAUSE Instrument malfunction. Improper slide placement.

ACTION TO TAKE Check that the interface cable and pipette locator are secure. Repeat the test in question, making sure that the slide is inserted correctly. If message recurs, call your Customer Support Center.

E14

RESULTS INVALID (Printout)

Sample/reference fluid levels are not adequate in dual sample cup. The DTE II Pipette is not working properly.

Assure that patient specimen and reference fluid volumes are adequate (at least 4 drops of each) in the dual sample cup. Verify pipette operation and proper protocol: - install the tips tightly. - check fluid levels in tips (both tips should have nearly equal fluid volumes). If fluid level is not even, dispense sample, eject tip, and insert new tips. - wipe outside of tips before dispensing fluid, being careful not to touch ends with tissue wipes (which will absorb fluid from tips). - 'keep pipette button depressed after spotting until you remove pipette from locator or fluid will be aspirated by pipette back off slide.

The pipette locator or boot is not seated correctly.

Remove pipette locator and nose assembly and make sure rubber boot is in place. Reposition pipette locator as it snaps into position. Be sure the DTE II Interface Cable (the cable that connects the DTE II Module to the back of the DT60 II System) is seated correctly.

The DTE II Interface Cable is not seated correctly.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

10-7

CODE/MESSAGE E15 RESULTS INVALID (Printout)

EXPLANATION/CAUSE Instrument malfunction. This message appears during option 43 along with the message: El 4 RESULTS INVALID. Instrument malfunction. This message appears during option 43 along with the message: E14 RESULTS INVALID. It may also appear when you are running a test.

ACTION TO TAKE Check the interface cable. If message recurs, call your Customer Support Center.

E16

RESULTS INVALID (Printout)

Check interface cable. If the message appears while you are running a test, run the test again. If message recurs, call your Customer Support Center. If message recurs at an excessive frequency, call your Customer Support Center.

E17

RESULTS INVALID (Printout)

Instrument malfunction. This message appears during option 43 along with the message: E14 RESULTS INVALID. Instrument malfunction.

E18

RESULTS INVALID (Printout)

Check the interface cable. Repeat the test in question. If message recurs at an excessive frequency, call your Customer Support Center.

E19

RESULTS INVALID (Printout)

Instrument malfunction.

Check the interface cable, the pipette and the pipette locator. Repeat the test in question. If message recurs at an excessive frequency, call your Customer Support Center.

10-8

Operator's Manual VITROS DT II System

5/95. Reprinted 1/99.

CODE/MESSAGE E20 A/D OUT OF RANGE OR BUSY

EXPLANATION/CAUSE Sample/reference fluid levels are not adequate in dual sample cup. The DTE II Pipette is not working properly.

ACTION TO TAKE Assure that patient specimen and reference fluid volumes are adequate (at least 4 drops of each) in the dual sample cup. Verify pipette operation and proper protocol: - install the tips tightly. - check fluid levels in tips (both tips should have nearly equal fluid volumes). If fluid level is not even, dispense sample, eject tip, and insert new tips. - wipe outside of tips before dispensing fluid, being careful not to touch ends with tissue wipes (which will absorb fluid from tips). - after spotting, keep the pipette button depressed until you remove the pipette from the locator or fluid will be aspirated off the slide. Remove pipette locator and nose assembly and make sure rubber boot is in place. Reposition pipette locator as it snaps into position. Be sure the DTE II Interface Cable (the cable that connects the DTE II Module to the back of the DT60 II System) is seated correctly. Discard any bottles open longer than the 30 day stability period.

The pipette locator or boot is not seated correctly.

The DTE II Interface Cable is not seated correctly.

Reference fluid is outdated.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

10-9

10.1.4 Instrument Function (F)


CODE/MESSAGE F12, F14, F16 DT 60 II SLIDE TRANSPORT ERROR (Printout) EXPLANATION/CAUSE A slide jam has been detected. ACTION TO TAKE Allow "tests in progress" to print out. Raise the pipette locator cover and check for slide jam. If there is a jam, remove the jammed slide. Check the pressure pad for binds by moving it up and down. Check the heated FORS weight for binds. To clear slides out of incubator, empty the slide disposal box and run option 3 (6 cycles) (VITROS DT60 II Chemistry System only). After the cycles have completed, check the slide disposal box. If the slide disposal box does not contain exactly one slide then a tracking error has occurred. Refer to section 8.3. Lower the pipette cover and resume testing when prompted by the analyzer. NOTE: When inserting slides into the DT60 II System, always insert the slide until the forward movement of the slide is stopped by the analyzer. F17 LINE VOLTAGE TOO LOW (Printout) Fluctuation in the internal power supply. Repeat tests that may have been deleted by the low line voltage.

10-10

Operator's Manual VITROS DT II System

Rev. 2004-03-30

CODE/MESSAGE F18 PRINTER MALFUNCTION (Printout)

EXPLANATION/CAUSE Paper jam occurred during loading.

ACTION TO TAKE Press PRINT on the keyboard. This may dislodge loose paper bits. Remove paper bits from top of printer. Turn the DT60 II System off and back on again. An easy way to load paper: - remove printer cover by pulling it up and towards the left, away from the analyzer. - cut the remaining paper away from core and pull the tape through top of print end. IMPORTANT: Do not pull tape backwards! This might damage paper feeding mechanism. - Cut corners of the fresh roll and position roll so that lead edge faces you and feeds from bottom of roll. Feed pointed end over roller and through slot in back of printer head until you see paper. Grab paper as it exits top of printer and pull a short distance through printer. Touch PRINT on keyboard overlay to advance paper. - Test printer by running option 4. NOTE: If the paper cannot be reloaded due to printer malfunction, test results may be obtained by pressing the TEST COMPLETE. To prevent this code, change the paper when a color strip appears.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

10-11

CODE/MESSAGE F19 MECHANISM ERROR (Printout)

EXPLANATION/CAUSE A slide jam has been detected.

ACTION TO TAKE Allow "tests in progress" to print out. Raise the pipette locator cover and check for slide jam. If there is a jam, remove the jammed slide. Check the pressure pad for binds by moving it up and down. Run option 3 for three cycles. Lower the pipette cover and resume testing when prompted by the analyzer.

F20

ELECTROM. POSITION ERROR (Display) DTSC TRANSFER MALFUNCTION (Printout)

DTE II Module malfunction.

Repeat the test. If message recurs, call your Customer Support Center. Remove the slide, if visible at the pickup station. Open the cover, if necessary, and remove the slide, then close the cover. Wait for the DTSC II Module to initialize (five minutes). Run option 108 and follow the instructions to step the slide through the slide transport cycle.

F30

A slide jam has been detected.

F31

SLIDE NOT AT READ STATION (Printout) SLIDE NOT AT BARCODE STATION (Printout)

Exit options and resume normal operation when prompted by the analyzer. NOTE: Does not require an operator response. Respond as instructed to the coded warning message that follows. To reduce slide jams, empty slide disposal box regularly.

F32

10-12

Operator's Manual VITROS DT II System

5/95. Reprinted 1/99.

CODE/MESSAGE F33 DTSC FILTER POSITION ERROR (Printout)

EXPLANATION/CAUSE DTSC II Module could not find home on the filter wheel, or is having problems with moving the f u . ., , & filter to the correct
p

ACTION TO TAKE Check to see that the arm is down at the read station, or make sure that a slide is at the read station. -,., n T c r , , , , u , Turn the DTSC IIu Module off and . .x . .i. c turn it on again waiting for it to reinitialize. Repeat tests that may have been deleted by turning the analyzer off. Clean the opal plug and sapphire window.

'

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

10-13

10.1.5 Temperature (H)


CODE/MESSAGE H11 TEMPERATURE MALFUNCTION (Display) ANALYZER TEMP. HIGH (Display) EXPLANATION/CAUSE Instrument malfunction. ACTION TO TAKE Call your Customer Support Center.

H12

Room temperature too high.

Check that the room temperature is within the range. The analyzer will not operate if the room temperature exceeds 29C (85F). If necessary, adjust the room thermostat. If the message appeared after the pipette locator cover was opened and closed, wait until the ANALYZER READY message is displayed. If neither of the first two probler..-, 3ms exists, call your Customer Support Center. Check that the room temperature is within the range. The analyzer will not operate if the room temperature falls below 15C(60F). If necessary, adjust the room thermostat. If the message appeared after the pipette locator cover was opened and closed, wait until the ANALYZER READY message is displayed. If the message appears when the analyzer is first turned on, wait until the ANALYZER READY message is displayed. If none of the first three problems exists, call your Customer Support Center.

Temporary temperature fluctuation after the pipette locator cover was open. Instrument malfunction.

HI 3

ANALYZER TEMP. LOW (Display)

Room temperature too low.

Temporary temperature decrease after the pipette locator cover was open.

Instrument is still warming up.

Instrument malfunction.

10-14

Operator's Manual V1TROS DT I! System

5/95. Reprinted 1/99.

CODE/MESSAGE H14 ANALYZER TEMP. LOW (Display)

EXPLANATION/CAUSE This is a warning that the room temperature is too low. This code will appear with another, more specific code. Test results printed on the DTE II Module with this message were for tests in progress when the module's internal temperature fluctuated outside its normal operating range. During continuous monitoring the DTSC II Module detects the preheat temperature is out of specifications.

ACTION TO TAKE Adjust the room temperature.

HI 5

TEMP MALFUNCTION (Printout)

H16

DTSC PREHEAT CAP TOO LOW/TOO HIGH THERMISTOR FAIL (Printout - only 1 response will print out) DTSC PREHEAT BOTTOM TOO LOW/ TOO HIGH THERMISTOR FAIL (Printout - only 1 response will print out) DTSC READ CAP TOO LOW/TOO HIGH THERMISTOR FAIL (Printout - only 1 response will print out)

Turn the DTSC II Module off then back on again, waiting for it to reinitialize. Repeat any tests that may have been deleted by turning the analyzer off. If the problem recurs, call your Customer Support Center. Adjust the room temperature.

H17

During temperature monitoring, the DTSC II Module detects that the preheat temperature is outside of specifications.

H18

During temperature monitoring, the DTSC II Module detects a problem with the temperature in the read station.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

10-15

CODE/MESSAGE H14-H19 Temperature Codes

EXPLANATION/CAUSE Inadequate space around the analyzer is preventing air circulation. Analyzer air exhaust vents dirty. A fan or vent is directed at the analyzer.

ACTION TO TAKE Allow air flow between the modules. Clean the vents with a soft-bristled brush after the instruments are powered off. Divert any direct air flow away from the analyzer. NOTE: If these actions fail to resolve the problem, power the instrument off and on.

10.1.6 Communications (N)


CODE/MESSAGE N11 TRANSMIT QUEUE FULL (Display) EXPLANATION/CAUSE The DT60 II System cannot queue any more messages to the DTSC II Module resulting in transmission problem. The DT60 II System cannot receive any more messages from the DTSC II Module. Problem with the processing of messages. The message that was sent from the DTSC II Module totheDT60 II System was undefined by the DT60 II System. ACTION TO TAKE Turn the DTSC II Module off then "power off" the DT60 II System. Turn the DT60 II System back on again then turn on the DTSC II Module, waiting for it to reinitialize. Repeat any tests that may have been deleted by turning the analyzer off. If the problem recurs, call your Customer Support Center.

N12

RECEIVER QUEUE FULL (Display)

N13

BAD RECEIVER COMMAND CODE (Printout)

10-16

Operator's Manual VITROS DT II System

5/95. Reprinted 1/99.

CODE/MESSAGE N14 BAD RECEIVER PROTOCOL (Printout)

EXPLANATION/CAUSE The DT60 II System did not receive the appropriate response from theDTSCII Module that it was expecting. The DT60 II System received a reading about a slide that it does not have defined. The DT60 II System received a reading that is out of sync with what was expected; either double readings or it missed a reading. There is a communications problem between the DT60 II System and DTSC II Module.

ACTION TO TAKE Turn the analyzer off then on again, waiting for it to reinitialize. Repeat any tests that may have been deleted by turning the analyzer off. If the problem recurs, call your Customer Support Center.

N15

BAD RATE SLIDE ID (Printout)

N16

DTSC READING OUT OF SYNC (Printout)

N17

SEQUENCE PROTOCOL ERROR (Printout)

Turn the DTSC II Module off then power off the DT60 II System. Turn the DT60 II System back on again, then turn on the DTSC II Module, waiting for it to reinitialize. Repeat any tests that may have been deleted by turning the analyzer off. If the problem recurs, call your Customer Support Center.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

10-17

10.1.7 Reflectometer (R) DT60II System


CODE/MESSAGE R11-R17 RESULTS INVALID (Printout) EXPLANATION/CAUSE The FORS head is dirty.

ACTION TO TAKE
Clean the FORS Head: lift the Spotting Station cover to expose the FORS weight. Lift up on the weight that covers the head. clean the FORS Head with a cotton swab moistened with water. Dry with a clean absorbent cloth before returning the weight into position over the FORS Head. Close the spotting station cover. Check for excessive room light. Light fixtures mounted directly above and in close proximity to the DT60 II System may affect the FORS. Make sure the slide disposal tray and main cover for the DT60 II System are in place. Repeatthe creatinine and ammonia slides to verify the test results. It is recommended that for optimum performance, creatinine and ammonia testing should not be performed when a slide for BUN is in the incubator.

The room light is excessive.

There is a light leak.

R18

PROTOCOL ERROR

You tried to run a slide for creatinine and ammonia immediately after you ran a slide for BUN and the result for BUN was > 40 mg/dL.

10-18

Operator's Manual VITROS DT II System

5/95. Reprinted 1/99.

10.1.8 Reflectometer (L) DTSC II Module


CODE/MESSAGE L11 INVALID RESULTS EXPLANATION/CAUSE ACTION TO TAKE

(Display)

The concentration of the sample is too high. The calibration is incorrect.

Dilute the sample using the appropriate diluent (refer to the Instructions for Use). If this code occurs with a high frequency and diluted samples produce normal results or another code, call your Customer Support Center. This Substrate Depletion code indicates that the change in kinetics for the test occurred too quickly.

L12, L13

INVALID RESULTS (Printout)

The sample has unusual kinetics, e.g., multiple myeloma. Calibration parameters were entered manually.

Dilute the sample using the appropriate diluent (refer to the Instructions for Use). Recalibrate. NOTE: Enzyme activity may be low, medium or high. These codes are designed to flag samples that contain interfering substances.

L14

RESULTS INVALID

There is some mechanical or electrical noise interfering with the calculation of the test result. A gain value sent from the DTSC II Module or a value that was computed for a new gain is out of the prescribed limits.

Rerun the test. If the problem recurs, call your Customer Support Center.

L15

GAIN OUT OF LIMITS FILTER CHANNEL (Printout)

Turn the analyzer off then clean the white reference plug and sapphire window. Turn the analyzer on again, waiting for the analyzer to reinitialize. If the problem recurs, call your Customer Support Center.

Rev. 2003-10-01

Operator's Manual VITROS DT II System

10-19

CODE/MESSAGE

EXPLANATION/CAUSE

ACTION TO TAKE

L16

MATH ERROR IN GAIN CALCULATION FILTER CHANNEL (Printout) GAIN READING OSCILLATING FILTER CHANNEL (Printout) INVALID DTSC SPOT (Printout)

The analyzer is unable to calculate new gain values.

Turn the analyzer off then on again, waiting for the analyzer to reinitialize. If the problem recurs, call your Customer Support Center. Turn the analyzer off then on again, waiting for the analyzer to reinitialize. If the problem recurs, call your Customer Support Center. If a slide is at the spotting station, and an invalid spot occurs, the DTSC II Module will invalidate the test and process the slide through the unit. If the slide is in the hold station (between the pickup and spotting station), the slide will be backed out to the pickup station. Insert a new slide and repeat the test. If fluid was dispensed at the spotting station without a slide present, clean the spotting station.

L17

Gain value is out of range, either above or below specification. The DTSC II Module detected a spot interrupt when unexpected. The slide was spotted too early before the green LED on the display was flashing, or there was no slide at the spotting station.

L18

L19

INVALID DTSC BARCODE (Printout)

Run Mode: Generation of slide loaded in the run mode has not yet been calibrated. Operator keyboard error (if number was entered manually). Calibration Mode: Incorrect calibrator kit for the test being calibrated. Remove the slide and calibrate the test. Remove the slide and enter correct gen. #. Use the correct calibrator kit:
Calibrator to Use
VITROS Specialty Calibrator Kit VITROS Isoenzyme Calibrator Kit VITROS Calibrator Kit

Test
*Theo & CHE *CKMB All Other

*C14 error code will occur on DT60 II Module. Calibration Data Module (CDM) does not contain the information required for the slide generation being used. Replace with the most current CDM. Install new CDMs when they arrive.

10-20

Operator's Manual VITROS DT II System

Rev. 2004-03-30

CODE/MESSAGE L20 L21 L22 L23 RATIO WHITE OUT OF RANGE (Printout) RATIO BLACK OUT OF RANGE (Printout) RATIO SAMPLE OUT OF RANGE (Printout) REFERENCE VOLTAGE OUT OF RANGE (Printout)

EXPLANATION/CAUSE The read area is dirty. The black offset reading has failed its limit check. The read area is dirty. The reference voltage reading has failed its limit check.-

ACTION TO TAKE Clean the opal plug and read window. See section 5 as a reference. Rerun the test. If the problem recurs, call your Customer Support Center.

Turn the DT60 II System off then on again, waiting for the analyzer to reinitialize. Repeat any tests that may have been deleted by turning the analyzer off. If the problem recurs, call your Customer Support Center. Check to see that the DTSC II Module cover is properly closed. The module will then reinitialize (5 minutes). Repeat any tests that may have been deleted by having the cover open. If the problem recurs, call your Customer Support Center

L27

DTSC COVER OPEN (Printout)

The cover to the DTSC II Module is open.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

10-21

10-22

Operator's Manual VITROS DT II System

5/95. Reprinted 1/99.

Installation and Site Specifications


The information contained in this section tells how to install the VITROS DT60 II Chemistry System, VITROS DTE II Module, and the VITROS DTSC II Module. This information will be primarily used by your Ortho-Clinical Diagnostics representative and your DT60 II System dealer when your equipment is initially installed. However, should you at some time need to move the analyzer and the module, this section will tell you how to prepare the equipment for a move, what to consider in selecting a new location for the equipment, and how to check the instrument's performance after it is reinstalled in a new site.

11.1 Installation
The DT60 II System will be unpacked and installed by an OrthoClinical Diagnostics representative or other trained personnel. CAUTION: Do not attempt to install a DT60 II System, DTE II Module, or a DTSC II Module unless properly trained. 1. Unpack equipment. Remove the DT60 II System, the DTE II Module, and the DTSC II Module from the shipping container. Remove packing materials from the: DT60 II System FORS Weight Pressure Pad Preheat Station DTE II Module Electrometer Nose Section Sample Holder DTSC II Module Operator Access Cover Preheat Heater Arm Read Station Heater Arm

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

11-1

2. Set up equipment. Place the DT60 II System, the DTE I! Module, and the DTSC II Module in the desired location. Position the DTE II Module to the right of the analyzer, and the DTSC II Module to the left. Do not plug in the analyzer yet. If the DTSC II Module is part of the installation then remove the connector shroud (Part No. 613871). Attach the DTE II Module cable assembly (Part No. 351572) to the rear of the DT60 II System chassis, securing it to the chassis with the screws provided. Connect the DTSC II Module to the adapter box using the DTE II Module's cable assembly(Part No. 338669) and secure it with the two connector screws provided. With the cover of the analyzer up, check to make sure that the 120/ 240 switch is in the proper position (120 position for the United States and Canada, 240 position for Europe). Replace the cover and secure the cable with the clamp (Part No. 338669). Insure that the line module/voltage selector is at the proper setting for the location. The setting may be changed by opening the snap-open cover of the module and rotating the selector drum to the proper setting. The voltage selected will appear in the window once the cover is closed. 3. Check power source and plug the analyzer in. Please refer to Electrical Requirements, 11.2.3. Check the line voltage. For the DT60 II System it must be between 105 and 127V ac. Check that the button of the analyzer is in the OFF position. Then, plug the analyzer into the receptacle. After the analyzer is plugged in, turn the DT60 II System on and wait while it goes through a series of self-checks and warms up (approximately 25 minutes). The DTSC II Module has settings for nominal voltages of 100, 120, 220, and 240V ac (-10% +5%). Check that the ON/OFF button on the module is in the OFF position. Then plug the module into the receptacle. After the module is plugged in, turn it on and wait while it goes through a series of self-checks and warms up (approximately 5 minutes). NOTE: The DT60 II System must be turned on first before the DTSC II Module. You do not need to wait for the analyzer to warm up before using the DTSC II Module.

11 -2

Operator's Manual VITROS DT II System

5/95. Reprinted 1/99.

4. Calibrate.
Calibrate the DT60 II System, DTE II Module, and DTSC II Module for all tests. 5. Run a quality control test. Run a quality control fluid for all tests to verify the calibration.

11.2 Site Specifications


11.2.1 Space Requirements
The analyzer and modules must rest upon a level bench or table large enough to accommodate the dimensions shown and sturdy enough to support the equipment's weight. Additional space should be provided adjacent to the analyzer, to permit easy operation and servicing. VITROS DT60 II System: Width Depth Height Weight = = = = 47.6cm (18.75 inches) 34.9cm (13.75 inches) 17.1cm (6.75 inches) Approximately 8.6kg (19 pounds)

VITROS DTE II Module: Width Depth Height Weight = = = = 14.6cm (5.75 inches) 35.4cm (13.9 inches) 16.5cm (6.5 inches) Approximately 2.7kg (6 pounds)

VITROS DTSC II Module: Width Depth Height Weight = 34.3cm (13.5 inches) = 35.4cm (13.9 inches) = 16.5cm (6.5 inches) = 7.7kg (1 7 pounds)

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

11 -3

Total space required, DT60 II System alone:

Width = 78.7cm (31 inches) Depth = 55.9cm (22 inches) Height = 48.3cm (19 inches) Total space required, DT60 II System and DTE II Module together: Width = 109.2cm (43 inches) Depth = 55.9cm (22 inches) Height = 48.3cm (19 inches) Total space required, DT60 II System and DTSC II Module together: Width = 127.8cm (50.3 inches) Depth = 55.9cm (22 inches) Height = 48.3cm (19 inches) Total space required, DT60 II System, DTE II Module, and the DTSC II Module together: Width = 157.5cm (62 inches) Depth = 55.9cm (22 inches) Height = 48.3cm (19 inches)

11.2.2 Environmental Requirements (Temperature, Humidity, and Altitude)


The DT60 II System, DTE II Module, and DTSC II Module are designed to operate effectively within" the temperature and humidity ranges typically found in physicians' offices. For effective operation, the temperature and relative humidity should be within the range indicated in the accompanying diagram. The figures below the diagram define the "corners" indicated in the diagram on the next page.

11-4

Operator's Manual VITROS DT II System

5/95. Reprinted 1/99.

Humidity

"1
58 133
:

I
| 66

]
; 68 |

1
70
!

I
i 72 ,

T
74

1
289

1""
30.0
:

:
:

60

62
;

64

144

15.6 ' 16.7

17.8 ' 18.9 ' 20.0 '' 21.1

22.2 ' 23.3

31.1

Temperature

15.5C(60F), 75% RH 15.5C(60F), 15%RH 29.4C (85F), 15% RH

29.4C (85F), 60% RH

(E) 23.9C(78F); 75% RH

11.2.3 Electrical Requirements


Both the DT60 II System and the DTSC II Module must be plugged into a properly grounded receptacle. Receptacle grounding should comply with all applicable electrical safety codes. Proper grounding is essential to assure proper operation and to minimize the likelihood of electrical damage to sensitive electronic circuits within the analyzer and modules. If you are uncertain about your electrical service, please consult a qualified electrician. The receptacle should be within 2.4 meters (8 feet) of the analyzer and DTSC II Module. The receptacle should be capable of supplying the following current: Nominal voltage 120 VAC 240 VAC DT60 II System 1 amp 0.5 amp DTSC II Module 0.5 amp 0.25 amp

Appliances (refrigerators, freezers, air conditioners, etc.) and other instruments can affect electrical power quality. These effects may be minimized by using a separate power circuit exclusively for the DT60 II System and DTSC II Module.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

11-5

11.2.4 Refrigerator and Freezer Space


Refrigerator and freezer space of 0.03 cubic meter (1 cubic foot) is required for storage of slides and test fluids.

11.3 Moving the Analyzer 11.3.1 Relocation Outside the Office


You may find it necessary to relocate the analyzer outside of your current office. If so, the new location must meet the same space, electrical, and environmental requirements as the original site. 1. Move the equipment carefully. Keep the equipment upright, and transport it in the same manner that you would transport any other type of office equipment. Before moving the equipment, make sure that no slides remain in the analyzer or module incubators, and turn the analyzer and DTSC II Module off. If the equipment is going to be transported a long distance, make sure that it is repacked and transported correctly. 2. Set up the equipment and run a quality control test. After the equipment is set up in the new location, a 30-minute environmental equilibration is required, followed by a quality control test for all relevant assays. When the results are within the acceptable range for your quality control system, the analyzer is ready to report results.

11.3.2 Relocation Within the Office


At some point, you may find it necessary to relocate the analyzer within your office after it has been installed. If you do so, the new location must meet the same space, electrical, and environmental requirements as the original site. 1. Move the equipment carefully. Before moving the equipment, make sure that there are no slides in the incubators of the DT60 II System, DTE II Module, or DTSC II Module. Turn the analyzer and the DTSC II Module OFF. Take care when moving the equipment since dropping or jarring could damage the equipment. 2. Set up the equipment and run a quality control test. After the equipment is set up in the new location, a 30-minute environmental equilibration is required, followed by a quality control test for all relevant assays. When the results are within the acceptable range for your quality control system, the analyzer is ready to report results.

11-6

Operator's Manual VITROS DT II System

5/95. Reprinted 1/99.

Warranty 12.1 New Equipment Warranty VITROS DT II System


1. Warranty Time Period Ortho-Clinical Diagnostics warrants the VITROS DT60 II System, VITROS DTE II Module, and VITROS DTSC II Module to function properly for one year from the date of initial installation, when installed within one year from date of shipment. This warranty covers the purchaser of this equipment and anyone else who owns it during the warranty period. 2. Warranty Repair Coverage If this equipment does not function properly during the warranty period, a Customer Technical Services representative will repair the equipment without charge during normal working hours (usually 8:00 a.m. to 5:00 p.m. Monday through Friday). Such repair service will include any adjustments and/or replacement of parts required to maintain your equipment in good working order. Supply items are billed as required. Off-hours service is available at overtime rates. 3. How to Obtain Services Call your Customer Support Center to obtain service. 4. Limitations Warranty service is limited to the contiguous United States. This warranty does not cover service or parts for any attachments, accessories or alterations not marketed by Ortho-Clinical Diagnostics, nor to correct problems resulting from their use. Ortho-Clinical Diagnostics makes no other warranties express, implied, or of merchantability for this equipment. Repair without charge is OCD's only obligation under this warranty. OCD will not be responsible for any consequential or incidental damages resulting from the sale, use, or improper functioning of this equipment, even if loss or damage is caused by the negligence or other fault of OCD. This limitation of liability will not apply to claims for injury to persons or damage to property caused by the sole negligence or fault of OCD.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

12-1

12.2 New Accessory Warranty VITROS DT Pipette and VITROS DTE Pipette
1. Warranty Time Period Ortho-Clinical Diagnostics warrants the VITROS DT Pipette and the VITROS DTE Pipette to function properly for one year from date of purchase. This warranty covers the purchaser of the pipette(s) and anyone else who owns it during the warranty period. 2. Warranty Repair Coverage If this equipment does not function properly during the warranty period, call your Customer Support Center. 3. Limitations Replacement without charge is OCD's only obligation under this warranty. Ortho-Clinical Diagnostics will not be responsible for any consequential or incidental damages resulting from the sale, use, or improper functioning of this equipment, even if loss or damage is caused by the negligence or other fault ofOCD. OCD makes no other warranties, express, implied, or of merchantability, for this equipment.

12-2

Operator's Manual VITROS DT II System

5/95. Reprinted 1/99.

CALIBRATION LOG
for VITROS DT II System
Date Calibrator Kit No. Reference Fluid Lot No. Operator. Exp. Date. Vitros DTE II Module _ Gen. No. Exp. Date. CDM No.. SLIDES
Test Lot Number Exp. Date

Serial Numbers: VitrosDT60 II System.

_ _

Pipette. Pipette.

Vitros DISC II Module.

CLM No. _ CALIBRATION COEFFICIENTS


CP#1 CP#2 CP#3 CP#4 DL#1* DL#3* QC Remarks

Affix dated Calibration Printout or manually record values in appropriate spaces. * DTSC il only

Make a photocopy of this log sheet prior to use for additional sheets.

Vitros is a trademark of Ortho-Clinical Diagnostics.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

13-1

SERVICE LOG
for VITROS DT II System

Vitros DT60 II System Serial Numbers: Vitros DTE II Serial Numbers: Vitros DTSC II Serial Numbers: Date

Analyzer. Module _ Module _ Problem Noted

Pipette _ Pipette. . Pipette. Corrective Action Taken Initial

Make a photocopy of this log sheet prior to use for additional sheets.

Vitros is a trademark of Ortho-Clinical Diagnostics.

13-2

Operator's Manual VITROS ' U System

5/95. Repr'

i 1/99.

TEST/REAGENT LOG
for VITROSDT II System

SLIDES Date Put Into Use Test


T T

REFERENCE FLUID Gen. No. Expiration Date Initials Date Opened Lot Number Gen. No. Exp. Initials Date

Lot Number

T T T T T
T

1 1

1 1 11
I 1 !
! 1

1 |

i
! 1 ! 1

1 I 1
1 I

! T LJ T
T

1 1 1 1 1

1 T LI T
T

T T T T T 1 |
1 1

1 ! 1 1 1

1 i 1 1

1 I 1
11

LJ T T

1 |

Make a photocopy of this log sheet prior to use for additional sheets.

Vitros is a trademark of Ortho-Clinical Diagnostics.

5/95. Reprinted 1/99.

Operator's Manual VITROS DT II System

13-3

VITROS DT II System Maintenance Log Maintenance Log


for the VITROS DT System
Daily Maintenance
Empty slide disposal box(es). Inspect pipettes and clean if necessary. Initials of person performing maintenance.

I Month

I Year

Refer to the "Instrument Care and Cleaning" section of your Operator's ManuaHor more details.
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 1819 20 21 22 23 24 25 26 27 28 29 30 31
I
I

Weekly Maintenance DT60


Clean the bar code reader and the drop detector surface. Clean pipette locator and visible slide track area. Monthly cleaning of the FORS head is recommended.

Weekly Maintenance DTE


Clean pipette locator and visible slide track area. Clean the rubber boot on the front of the electrometer.

Weekly Maintenance DTSC


Clean the pickup and slide spotting stations. Clean the pipette locator. Clean the slide track. Clean the reference cap and the sapphire read window. Initials of person performing maintenance.

Supplies Inventory
Check expiration dates and inventory of the following supplies: Slides Reference fluid Calibrators Controls Printer paper

Points to consider: Do you have enought for the next month? for the next 6 months? Do you have more than one lot number for any chemistry? Store new lot numbers separately. Are supplies being stored at the correct temperature?

stora e 9 Temperature Guide -

CAT

L o tN a

"

1 1
JIL
Lo

Sic e Of 1So> 0

Before using this page, please make copies for future use

13-4

Operator's Manual VITROS DT II System

5/95. Reprinted 1/99.

VITROS DT II System Quality Control Log


CONTROL Lot No.
Note: When the control lot no. changes, use the new control assay sheet to find acceptable ranges and begin a new control log sheet.

Test name

Range Acceptable? Corrective Action/Comments Initials

H|HH| ^HHi
^H ^H

Date

Control Value

H H H HHH^HHHHIH^^H ^B

^HiH BHHHHIH HHH^HIHHHHHH ^B ^^H^^^^^^HBBI^H HI ^^^^^IBHHI^^H^^^^H^H^^^HI I^HHII^^HIHI^^^HHH^I HH I^HH B^^HHI^i^^HHi^H BB HHIH IHIH ^H IHHH I^HII HUH ^HI^^^^^^HH^^^S H I HHIH ^H IHHH I^HHI ^ ^ ^ ^ H H ^ ^ ^ ^ ^ ^ ^ ^ ^ B ^; _^^^^_^_ HI HHIH HH^H IHI^^^HHI^I^^^HH IB HHIHII^HHI ^^^HIHIi^^B^^H^H ^H HHUH ^HI^^H^^^^^^H^H ^H ^H HH^^B II^^HI i^^^^^^HHHH^^H ^H BH ^i^iH HH^^^^HHHHHIH HiHH 1
Before using this page, please make copies for future use. Operator's Manual VITROS DT II System 13-5 5/95. Reprinted 1/99.

HHIHHHI i^^HH ^H ^HHH ^^BH^H^^HHBHI ^B IHBB I^^HH HHHHHHIHHHH ^B ^H

Levey-Jennings Quality Control Chart for VITROS DT II System

Date. Slide Lot #.

Gen#_ Control. Control Lot #.

Level 1 Date +2SD = +1 S D = Mean = -1 S D = -2SD = Observed T . , . , . . . . . . . . . . . . . . . . . . . . . . . . . . . V a u / / / / / / / / / / / / / / / / / / / / / / / / / / / / / / / l e


Control Lot #.

Level 2

Date (_ +2 SD = +1 SD = Mean =
-

\ '

-1 SD = -2SD =

Observed I I Value / / . / / / . / / / / / / / . / / . / / / / / / / / / . / / / / / / / /
Corrective Action:

ii

Reviewed: Date

Signature.

13-6

Operator's Manual VITROS DT II System

5/95. Reprinted 1/99.

Coronary Risk Classification Questionnaire


Please answer the following questions. This information will be used with the Vilros DT60 II Chemistry System. Name: Female Male Yes Age No Today's Date Date of Birth: / / / /_

Do you smoke or have you quit smoking within the last 12 months? Are you diabetic? Yes No

Do you have a history of cardiovascular disease (left-ventricular hypertrophy)? For internal use only Systolic Blood Pressure Notes: CHOL

Yes

No

Not sure

HDLC

I f Ortho-Clinical Diagnostics
fif4fc company

Coronary Risk Classification Questionnaire


Please answer the following questions. This information will be used with the Vitros DT60 II Chemistry System. Name: Female Male Yes Age No Today's Date Date of Birth: / / / /_

Do you smoke or have you quit smoking within the last 12 months? Are you diabetic? Yes No

Do you have a history of cardiovascular disease (left-ventricular hypertrophy)? For internal use only Systolic Blortd Pressure Notes: CHOL

Yes

No

Not sure

HDLC

' Ortho-Clinical Diagnostics


company

Coronary Risk Classification Questionnaire


Please answer the following questions. This information will be used with the Vitros DT60 II Chemistry System. Name: Female Male Yes Age No Today's Date Date of Birth: / / / /_

Do you smoke or have you quit smoking within the last 12 months? Are you diabetic? Yes No

Do you have a history of cardiovascular disease (left-ventricular hypertrophy)? For internal use only Systolic Blood Pressure Notes: CHOL

Yes

No

Not sure

HDLC

' Ortho:Clinical Diagnostics


company

13-8

Operator's Manual VITROS DT II System

5/95. Reprinted 1/99.

C hem

i stry

TABLE OF CONTENTS-DT SLIDES


Instructions For Use
Abbreviation ALBDT ALKP DT ALTDT AWIYL DT ASTDT BUN/UREA DT CaDT CHEDT CHOL DT CKDT CKMB DT CIDT CO 2 DT CREA DT CRSC DT FeDT GGTDT GLUDT HDLC DT HDLC DT Test Name Albumin Alkaline Phosphatase Alanine Aminotransferase Amylase Aspartate Aminotransferase Urea Nitrogen Calcium Cholinesterase Cholesterol Creatine Kinase Creatine Kinase MB Chloride Carbon Dioxide Creatinine (Blank-Corrected Method) Creatinine (Single-Slide Method) Iron Glutamyltransferase Glucose HDL Cholesterol (Using the VITROS DT HDL Cholesterol Kit) HDL Cholesterol (Using the VITROS DT Micro HDL Cholesterol Kit) Potassium Lactate Lactate Dehydrogenase Lithium Lipase Magnesium Sodium Neonatal Bilirubin Ammonia Phosphorus
Operator's Manual VITROS DT II System

Pub.

No.

Version 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0


1.0

Date 2003-10-01 2003-04-30 2003-10-01 2003-08-11 2003-10-01 2003-10-01 2003-03-28 2003-10-01 2003-03-28 2003-10-01 2003-04-30 2003-10-01 2003-08-11 2003-10-01 2003-04-30 2003-10-01 2003-08-11 2003-08-11 2004-02-29 2004-02-29

C-355 C-337 C-336 C-311 C-338 C-301 C-348 C-358 C-304 C-342 C-351 C-309 C-308 C-334 C-353 C-371 C-343 C-300 C-341_EN C-354_EN

1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 2.0 2.0

K + DT LACDT LDHDT LiDT LIPA DT MgDT Na DT NBIL DT NH3 DT (AMON) DT PHOS DT


Rev. 2004-03-30
+

C-306 C-357 C-344 C-372 C-356 C-349 C-307 C-364 C-333 C-350

1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0

2003-03-28 2003-10-01 2003-10-01 2003-08-11 2003-04-30 2003-10-01 2003-10-01 2003-10-01 2003-10-01 2003-04-30
14-1

TABIE OF CONTENTS--DT SHOES


Instructions for Use

Abbreviation TBIL DT THEO DT TPDT TRIG DT urCR DT URIC DT

Test Name Total Bilirubin Theophylline Total Protein Triglycerides Urine Creatinine (Single-Slide Method) Uric Acid

Pub. No. C-305 C-347 C-310 C-303 C-373 C-302

Version 1.0 1.0 1.0 1.0 1.0 1.0

Date 2003-10-01 2003-10-01 2003-10-01 2003-04-30 2003-10-01 2003-08-11

DT Calibrator DT Isoenzyme Calibrator DT Specialty DT Control DT Isoenzyme Control DT Reference Fluid 7% BSA

DT Calibrator DT Isoenzyme Calibrator Kit DT Specialty Calibrator Kit DT Control DT Isoenzyme Control DT Reference Fluid 7% BSA

J23110_EN J23112_EN J23115_EN J23111_EN J23113_EN J23114_EN J11460

3.0 2.0 2.0 2.0 2.0 2.0 2.0

2004-03-31 2004-02-29 2004-02-29 2004-02-29 2004-02-29 2004-02-29 2003-07-28

14-2

Operator's Manual VITROS DT II System

Rev. 2004-03-30

Chemistry

INSTRUCTIONS FOR USE


VITROS Chemistry Products ALB DT Slides Intended Use
For in vitro diagnostic use only. VITROS ALB DT Slides quantitatively measure albumin (ALB) concentration in serum and plasma.

AIRDT
Albumin

Summary and Explanation of the Test


Of all serum proteins, albumin is present in the highest concentration. It maintains the plasma oncotic pressure and the transport of many substances. Increased serum albumin may indicate dehydration or hyperinfusion with albumin; a decrease is found in rapid hydration, overhydration, severe malnutrition and malabsorption, severe diffuse liver necrosis, chronic active hepatitis, and neoplasia. Albumin is commonly reduced in chronic alcoholism, pregnancy, renal protein loss, thyroid dysfunction, peptic ulcer disease, and chronic inflammatory diseases.1

Principles of the Procedure


The VITROS ALB DT Slide method is performed using the VITROS ALB DT Slide and the VITROS Chemistry Products DT Calibrator Kit on VITROS DT60 II Chemistry Systems. The VITROS ALB DT Slide is a multilayered, analytical element coated on a polyester support. A drop of patient sample is deposited on the slide and is evenly distributed by the spreading layer to the underlying layers. When the fluid penetrates the reagent layer, the bromcresol green (BCG) dye diffuses to the spreading layer and binds to albumin from the sample. This binding results in a shift in wavelength of the reflectance maximum of the free dye. The color complex that forms is measured by reflectance spectrophotometry. The amount of albumin-bound dye is proportional to the concentration of albumin in the sample.

Reaction Sequence
albumin + bromcresol green (BCG) BCG-albumin complex

Test Type and Conditions


Test Type and Conditions for ALB DT
Test Type Colorimetric VITROS DT60 II Module DTSC II Approximate Incubation Time 3 minutes Temperature 37C (98.6F) Wavelength 630 nm Sample Drop Volume 10 uL

Warnings and Precautions


For in vitro diagnostic use only. Take care when handling materials and samples of human origin. Since no test method can offer complete assurance that infectious agents are absent, consider all clinical specimens, controls, and calibrators potentially infectious. Handle specimens, solid and liquid waste, and test components in accordance with local regulations and NCCLS Guideline M292 or other published biohazard safety guidelines. For specific warnings and precautions for calibrators, quality control materials, and other components, refer to the instructions for use for the appropriate VITROS product, or to other manufacturer's product literature.

Version 1.0

Pub.

No. C-355

AlBDT
Albumin

INSTRUCTIONS FOR USE


Reagents

Reagents
Slide Diagram

Slide Ingredients
Reactive ingredients per cm
2

Bromcresol green dye 104 ng.

Other ingredients
Polymer beads, binders, buffer, and surfactants.

1. Upper slide mount 2. Spreading layer (beads) 3. Reagent layer bromcresol green dye buffer, pH 3.1 4. Support layer 5. Lower slide mount

Slide Handling
Do not use slides with damaged or incompletely sealed packaging.

Slide Preparation
IMPORTANT: The slide must reach room temperature, 18-28 C (64 -82 F), before the wrapper is opened. Do not use unopened slides that have been at room temperature, 18"-28 X: (64 -82 F) for >48 hours. 1. 2. Remove the unopened slide from the box. \Narm the unopened slide for 15 minutes. Unopened slides that remain in the sealed wrapper may be returned to refrigerator or freezer storage. 3. Remove the packaging and place the slide into the loading station within 15 minutes after opening.

Slide Storage and Stability


VITROS ALB DT Slides are stable until the expiration date on the carton when they are stored and handled as specified.

Slide Storage and Stability for ALB DT


Slides Unopened

Opened

Storage Condition Room temperature 18-28C (64-82F) Refrigerated 2-8C (36-46F) Frozen <-18C (<0F) Room temperature 18-28C (64-82F)

Stability <48 hours Until expiration date Until expiration date <15 minutes

Specimen Requirements
Handle specimens as biohazardous material.

Specimens Recommended
Serum Plasma:3 Heparin Certain collection devices have been reported to affect other analytes and tests.4 Confirm that your collection devices are compatible with this test.

IMPORTANT:

Serum and Plasma


Specimen Collection and Preparation
Collect specimens using standard laboratory procedures. 56 Patient Preparation No special patient preparation is necessary. Special Precautions For the effect of hemolysis on test results, refer to "Limitations of the Procedure." Albumin concentrations vary with posture. Results from an upright posture may be approximately 0.3 g/dL (3 g/L) higher than those from a recumbent posture.7 Centrifuge specimens and remove the serum or plasma from the cellular material within 3 days of collection.8 Pub. No. C-355 Version 1.0

INSTRUCTIONS FOR USE


Testing Procedure Specimen Handling and Storage Handle and store specimens in stoppered containers to avoid contamination and evaporation. Mix samples by gentle inversion and bring to room temperature, 18-28C (64-82F), prior to analysis. Specimen Storage and Stability for ALB DT: Serum Temperature Storage 18-28C(64-82F) Room temperature 2-8C (36-46F) Refrigerated <-18C(<0F) Frozen and Plasma 8 Stability <7 days <1 month Indefinite

AIR DT
Albumin

Testing Procedure
Materials Provided
VITROS Chemistry Products ALB DT Slides

Materials Required But Not Provided


| VITROS Chemistry Products DT Calibrator Kit Quality control materials, such as VITROS Chemistry Products DT Control I & II Isotonic saline or reagent-grade water VITROS DT Pipette

Operating Instructions
Refer to the operator's manual for your VITROS DT60 II Chemistry System. PANT; Bring all fluids and samples to room temperature, 18-28C (64-82F), prior to analysis.

Sample Dilution
| If albumin concentrations exceed the system's reportable (dynamic) range: 1. Dilute the sample with one part isotonic saline or reagent-grade water to 10 parts sample. 2. Reanalyze. 3. Multiply the results by 1.1 to obtain an estimate of the original sample's albumin concentration.

Calibration
Required Calibrators
VITROS Chemistry Products DT Calibrator Kit, bottles 1, 2, and 4

Calibrator Preparation, Handling, and Storage


Refer to the instructions for use for VITROS DT Calibrator Kit.

Calibration Procedure
Refer to the operator's manual for your VITROS DT60 II Chemistry System.

When to Calibrate
Calibrate: When the slide lot number changes. When critical system parts are replaced due to service or maintenance. When government regulations require. - For example, in the USA, CLIA regulations require calibration or calibration verification at least once every six months. The VITROS ALB DT test may also need to be calibrated: If quality control results are consistently outside acceptable range. After certain service procedures have been performed. For additional information, refer to the operator's manual for your VITROS DT60 II Chemistry System.

Calculations
Reflectance from the slide is measured at 630 nm after the fixed incubation time. Once a calibration has been performed for each slide lot, albumin concentration in unknown samples can be determined using the software-resident endpoint colorimetric math model and the response obtained from each unknown test slide.

Version 1.0

Pub. No. C-355

ALBDT
Albumin

INSTRUCTIONS FOR USE


Quality Control
Calibration parameters are automatically assessed by the VITROS DT60 II Chemistry System against a set of quality parameters resident within the analyzer software. Failure to meet any of the pre-defined quality parameters results in a failed calibration. The calibration report should be used in conjunction with quality control results to determine the validity of a calibration.

Validity of a Calibration

Reportable (Dynamic) Range Reportable (Dynamic) Range for ALB DT


Conventional (g/dL) 1.0-6.0 SI Units (g/L) 10-60

For out-of-range samples, refer to "Sample Dilution."

Traceability of the Calibration


Values assigned to the VITROS Chemistry Products DT Calibrator Kit for albumin are traceable to the Certified NIST (National Institute of Standards and Technology) Total Protein Standard Reference Material, SRM* (Standard Reference Material) 927c. The Ortho-Clinical Diagnostics calibration laboratory uses SRM" 927c to assign values to a series of working human albumin standards prepared from purified human albumin. The working standard series is used to calibrate a bromcresol green albumin method9 to support value assignment for the VITROS DT Calibrator Kit.

Quality Control
Procedure Recommendations
| WARNING: Handle quality control materials as biohazardous material. Choose control levels that check the clinically relevant range. Analyze quality control materials in the same manner as patient samples, before or during patient sample processing. To verify system performance, analyze control materials: - After calibration. - According to local regulations or at least once each day that the test is being performed. - After specified service procedures are performed. Refer to the operator's manual for your VITROS DT60 II System. If control results fall outside your acceptable range, investigate the cause before deciding whether to report patient results. For general quality control recommendations, refer to Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition10 or other published guidelines. For additional information, refer to the operator's manual for your VITROS DT60II Chemistry System.

Quality Control Material Selection

I
I I

MPORTANT;

VITROS DT Control I & II are recommended for use with the VITROS DT60 II Chemistry System. Evaluate the performance of other commercial control fluids for compatibility with this test before using for quality control.

Control materials other than VITROS DT Controls I & II may show a difference when compared with other albumin methods if they: - Depart from a true h uman matrix. - Contain high concentrations of preservatives, stabilizers, or other nonphysiological additives. Controls low in carbon dioxide concentration may show a negative bias11 that can be avoided by reconstituting lyophilates with a bicarbonate diluent instead of water. Do not use control materials stabilized with ethylene glycol.

Quality Control Material Preparation and Storage


Refer to the instructions for use for VITROS Chemistry Products DT Control I & II or to other manufacturer's product literature.

Pub. No. C-355

Version 1.0

[3 VMTFJCpS

INSTRUCTIONS FOR USE


Expected Values and Reporting Units

ALBDT
Albumin

Expected Values and Reporting Units


Reference Interval Reference Interval for ALB DT12 Conventional Units (g/dL) 3.5-5.0

SI Units (g/L) 35-50

Each laboratory should confirm the validity of these intervals for the population it serves.

Reporting Units and Unit Conversion


The VITROS DT60 II Chemistry System may be programmed to report albumin results in conventional and SI units. Reporting Units and Unit Conversion for ALB DT Conventional Units SI Units g/dL g/L (g/dL x 10)

Limitations of the Procedure


Known Interferences
The VITROS ALB DT Slide method was screened for interfering substances following NCCLS Protocol EP7.13 The substances listed in the table, when tested at the concentrations indicated, caused the bias shown. Known Interfering Substances for ALB DT Interferent Concentration Interferent* Conv. (mg/dL] Si (g/L) Hemoglobin
100

Albumin Concentration Conv. (g/dL) SI(g/L)


3.5 35

Bias Conventional
6%

SI
6%

200 400
*

(1.0) (2.0) (4.0)

3.5 3.5

35 35

13% 26%

13% 26%

It is possible that other interfering substances may be encountered. These results are representative; however, your results may differ somewhat due to test-to-test variation. The degree of interference at concentrations other than those listed might not be predictable.

Other Limitations
Certain drugs and clinical conditions are known to alter albumin concentrations in vivo. For additional information, refer to one of the published summaries.141S

Version 1.0

Pub. No. C-355

AlBDT
Albumin

INSTRUCTIONS FOR USE


Performance Characteristics

Performance Characteristics
Method Comparison

The plots and table show the results of a comparison of samples analyzed on the VITROS DT60 II System with those analyzed using the bromcresol green dye-binding comparative method.11

Method Comparison for ALB DT: Serum Conventional Units


70605040
Q W O

SI Units

y =x

30 20 > 100 0 10 20 30 40 50 60 70

Comparative Method: Bromcresol Green (9/dL)

Comparative Method: Bromcresol Green (gfl.)

Method Comparison for ALB DT: Serum


Conventional Units (g/dL) n DT60 II System vs. comparative method 79 Correlation Slope Coefficient 0.98 0.992 Range of Sample Cone. 1.4-5.8 Intercept Sy.x 0.06 0.14 SI Units (g/L) Range of Sample Cone. Intercept 0.59 Sy.x 1.44

14-58

Precision
Precision was evaluated with quality control materials on the VITROS DT60 II System following NCCLS Protocol EP5.16 The data presented are a representation of test performance and are provided as a guideline. Variables such as sample handling and storage, reagent handling and storage, laboratory environment, and system maintenance can affect reproducibility of test results.

Precision for ALB DT: Serum


Conventional Units <g>dL) Within Within Mean Cone. Day SD* Lab SD** 2.5 0.02 0.06 4.4 0.04 0.10 SI Units (g/L) Mean Cone. 25 44 Within Day SD* 0.2 0.4 Within Lab SD** 0.6 1.0 Within Lab CV%** 2.6 2.3 No. Observ. 88 88 No. Days 22 22

System VITROS DT60 II

Within Day precision was determined using two runs/day with two replications. Within Lab precision was determined using a single lot of slides and calibrating weekly.

Pub. No. C-355

Version 1.0

INSTRUCTIONS FOR USE


References

AlBDT
Albumin

References
1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16. Tietz NW (ed). Fundamentals of Clinical Chemistry, ed. 3. Philadelphia: WB Saunders; 328-329; 1987. NCCLS. Protection of Laboratory Workers from Instrument Biohazards and Infectious Diseases Transmitted by Blood, Body Fluids and Tissue; Approved Guideline. NCCLS Document M29 (OSBN 1-56238). NCCLS, Wayne, PA 19087; 1997. Doumas BT, et al. Differences Between Values for Plasma and Serum in Tests Performed in the Ektachem 700 XR Analyzer, and Evaluation of "Plasma Separator Tubes (PST)." Clin. Chem. 35:151; 1989. Calam RR. Specimen Processing Separator Gels: An Update. J Clin Immunoassay. 11:86-90; 1988. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Venipuncture. NCCLS Document H3. Wayne, PA: NCCLS; 1991. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Skin Puncture. NCCLS Document H4. Wayne, PA: NCCLS; 1991. Tietz NW. Textbook of Clinical Chemistry. Philadelphia: WB Saunders; 589; 1986. Clinical Laboratory Handbook for Patient Preparation and Specimen Handling. Fascicle VI: Chemistry/Clinical Microscopy. Northfield, IL: College of American Pathologists; 1992. Doumas BT, Biggs HG. Determination of serum albumin. Standard Methods in Clinical Chemistry. 7:175-188; 1972. NCCLS. Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition. NCCLS Document C24. Wayne, PA: NCCLS; 1999. Corcoran RM, Durnan SM. Albumin Determination by a Modified Bromcresol Green Method. Clin. Chem. 23(4):765; 1977. Peters T. All about Albumin. San Diego: Academic Press; 256; 1996. NCCLS. Interference Testing in Clinical Chemistry. NCCLS Document EP7. Wayne, PA: NCCLS; 1986. Young DS. Effects of Drugs on Clinical Laboratory Tests, ed. 4. Washington D.C.: AACC Press; 1995. Friedman RB. Young DS. Effects of Disease on Clinical Laboratory Tests. Washington, D.C.: AACC Press; 1990. NCCLS. User Evaluation of Precision Performance with Clinical Chemistry Devices. NCCLS Document EP5. Wayne, PA: NCCLS;

1992.

Glossary of Symbols
The Glossary of Symbols table defines the symbols used on the VITROS Chemistry Products packaging and on the VITROS Chemistry Products Assay Sheets.

Glossary of Symbols
Do Not Reuse Use by or Expiration Date (YYYY-MM-DD) Lot Number Manufacturer's Serial Number Catalog Number or Product Code Attention: See Instructions for Use. Manufacturer Authorized Representative Contains Sufficient for "n" Tests For In Vitro Diagnostic Use Store At or Below Store Between Consult Instructions for Use

Fragile, Handle with Care. Keep Dry This end up

Store At or Above

Version 1.0

Pub. No. C-355

VITR_[fi5 0

ALBDT
Albumin

INSTRUCTIONS FOR USE


Revision History

Revision History
Date of Revision 2003-10-01 Version 1.0 Description of Technical Changes* New format New organization and sections consistent with IVD Directive Specimen Storage and Stability - updated stability values Materials Required But Not Provided and Sample Dilution - added reagent-grade water Quality Control Material Selection - added statements regarding controls low in carbon dioxide concentration and ethylene glycol Known Interferences - updated values Method Comparison - updated all data and the plot Precision - updated all data References - added all except 7

The change bars indicate the position of a technical amendment to the text with respect to the previous version of the document.

When this Instructions For Use is replaced, sign and date below and retain as specified by local regulations or laboratory policies, as appropriate.

Signature

Obsolete Date

C
Ortho-Clinical Diagnostics Mandeville House 62 The Broadway Amersham Buckinghamshire HP7 OHJ England Ortho-Clinical Diagnostics, Inc. 100 Indigo Creek Drive Rochester, NY 14626-5101

' Ortho-Clinical Diagnostics


a ^e/hmtm^fotmmn company

VITROS is a trademark of Ortho-Clinical Diagnostics, Inc. Ortho-Clinical Diagnostics, Inc., 2003. All rights reserved. Printed in USA.

Pub. No. C-355

Version 1.0

VITR

Chemistry

INSTRUCTIONS FOR USE


VITROS Chemistry Products ALKP DT Slides Intended Use

ALKP OT
Alkaline Phosphatase

For in vitro diagnostic use only. VITROS ALKP DT Slides quantitatively measure alkaline phosphatase (ALKP) activity in serum and plasma.

Summary and Explanation of the Test


Alkaline phosphatase is present mainly in bone, liver, kidney, intestine, placenta, and lung. Serum alkaline phosphatase may be elevated in increased bone metabolism, for example, in adolescents and during the healing of a fracture; primary and secondary hyperparathyroidism; Paget's disease of bone; carcinoma metastatic to bone; osteogenic sarcoma; and Hodgkin's disease if bones are invaded. Hepatobiliary diseases involving cholestasis, inflammation, or cirrhosis increase alkaline phosphatase activity; alkaline phosphatase activity may be increased in renal infarction and failure and in the complications of pregnancy. Low alkaline phosphatase activity may occasionally be seen in hypothyroidism.1

Principles of the Procedure


The VITROS ALKP DT Slide method is performed using the VITROS ALKP DT Slide and the VITROS Chemistry Products DT Calibrator Kit on VITROS DT60/DT60 II Chemistry Systems. The VITROS ALKP DT Slide is a multilayered, analytical element coated on a polyester support. A drop of patient sample is deposited on the slide and is evenly distributed by the spreading layer to the underlying layers. The spreading layer contains the p-nitrophenyl phosphate substrate and other components needed for the reaction. The ALKP in the sample catalyzes the hydrolysis of the p-nitrophenyl phosphate to p-nitrophenol at alkaline pH. The p-nitrophenol diffuses into the underlying layer, and it is monitored by reflectance spectrophotometry. The rate of change in reflection density is converted to enzyme activity.

Reaction Sequence
p-nitrophenyl phosphate
ALKP Mg ,AMP

p-nitrophenol + H3PO4

Test Type and Conditions


Test Type and Conditions for ALKP DT
Test Type Rate VITROS DT60/DT60 II Module DTSC/DTSC II Approximate Incubation Time 5 minutes Temperature 37C (98.6F) Wavelength 400 nm Sample Drop Volume 10 |JL

Warnings and Precautions


For in vitro diagnostic use only. Take care when handling materials and samples of human origin. Since no test method can offer complete assurance that infectious agents are absent, consider all clinical specimens, controls, and calibrators potentially infectious. Handle specimens, solid and liquid waste, and test components in accordance with local regulations and NCCLS Guideline M292 or other published biohazard safety guidelines. For specific warnings and precautions for calibrators, quality control materials, and other components, refer to the instructions for use for the appropriate VITROS product, or to other manufacturer's product literature.

Version 1.0

Pub.

No. C-337

VITRI

A1KPDT
Alkaline Phosphatase

INSTRUCTIONS FOR USE


Reagents

Reagents
Slide Diagram

Slide Ingredients
Reactive ingredients per cm2
p-nitrophenyl phosphate 55 ug; 2-amino-2-methyl-1-propanol (AMP) 0.1 mg; and magnesium sulfate 1.6 ug.
,-'

i - 2 , 3

Other ingredients
Pigment, binders, buffers, surfactants, cross-linking agent and stabilizer.

" ~ - -.

1. Upper slide mount 2. Spreading layer (BaSO4) magnesium sulfate AMP p-nitrophenyl phosphate 3. Reagent layer buffer, pH 10.5 4. Support layer 6. Lower slide mount

Slide Handling
Do not use slides with damaged or incompletely sealed packaging.

Slide Preparation
IMPORTANT: The slide must reach room temperature, 18-28C (64-82F), before the wrapper is opened. Do not use unopened slides that have been at room temperature, 18-28C (64-82F) for >48 hours. 1. 2. Remove the individual slides from the box. Warm the unopened slide for at least 15 minutes. Unopened slides that remain in the sealed wrapper may be returned to refrigerator or freezer storage. 3. Remove the packaging and place the slide into the loading station within 15 minutes after opening.

Slide Storage and Stability


VITROS ALKP DT Slides are stable until the expiration date on the carton when they are stored and handled as specified.

Slide Storage and Stability for ALKP DT Slides Storage Condition


Unopened Opened Room temperature Refrigerated Frozen Room temperature 18-28C (64-82F) 2-8C (36-46F) <-18C (<0F) 18-28C (64-82F)

Stability <48 hours Until expiration date Until expiration date <15 minutes

Specimen Requirements
Handle specimens as biohazardous material.

Specimens Recommended

Serum Plasma:3 IMPORTANT;

Heparin Certain collection devices have been reported to affect other analytes and tests4 Confirm that your collection devices are compatible with this test.

Specimens Not Recommended


Plasma:5 EDTA Citrate Fluoride oxalate Do not use hemolyzed specimens.

Pub. No. C-337

Version 1.0

|S] VITRJIS

INSTRUCTIONS FOR USE


Testing Procedure Serum and Plasma Specimen Collection and Preparation
Collect specimens using standard laboratory procedures.6 7 Patient Preparation No special patient preparation is necessary. Special Precautions For the affect of high concentration of bilirubin on test results, refer to "Limitations of the Procedure." Centrifuge specimens and remove the serum from the cellular material within 4 hours of collection.5

ALKPDT
Alkaline Phosphatase

Specimen Handling and Storage


Handle specimens as biohazardous material. Handle and store specimens in stoppered containers to avoid contamination and evaporation. Mix samples by gentle inversion and bring to room temperature, 18-28C (64-82F), prior to analysis.

Specimen Storage and Stability for ALKP DT: Serum and Plasma5 Storage Temperature Stability
Room temperature Refrigerated Frozen 18-28OC(64-82OF) 2-8C (36-46F) <-18C(<0F) <4 days <4 days <4 days

Testing Procedure
Materials Provided
VITROS Chemistry Products ALKP DT Slides

Materials Required But Not Provided


VITROS Chemistry Products DT Calibrator Kit Quality control materials, such as VITROS Chemistry Products DT Control I & II VITROS Chemistry Products 7% BSA VITROS DT Pipette Isotonic saline

Operating Instructions
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System. Bring all fluids and samples to room temperature, 18-28C (64-82F), prior to analysis. iMPORTANT:

Sample Dilution
| If alkaline phosphatase activities exceed the system's reportable (dynamic) range: 1. Dilute with isotonic saline or VITROS 7% BSA. 2. Reanalyze. 3. Multiply the results by the dilution factor to obtain an estimate of the original sample's alkaline phosphatase activity.

Calibration
Required Calibrators
VITROS Chemistry Products DT Calibrator Kit, bottles 1, 2, and 4

Calibrator Preparation, Handling, and Storage


Refer to the instructions for use for VITROS DT Calibrator Kit.

Calibration Procedure
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Version 1.0

Pub. No. C-337

VI"TFj[C|3"S 0

AIKPDT
Alkaline Phosphatase

INSTRUCTIONS FOR USE


Quality Control

When to Calibrate
Calibrate: When the slide lot number changes. When critical system parts are replaced due to service or maintenance. When government regulations require. - For example, in the USA, CLIA regulations require calibration or calibration verification at least once every six months. The VITROS ALKP DT test may also need to be calibrated: If quality control results are consistently outside acceptable range. After certain service procedures have been performed. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Calculations
Based on sequential readings of the slide's reflectance at 400 nm over the defined incubation period, a rate of change in reflectance is determined. This rate is used in the software-resident multi-point rate calibration model to compute enzyme activity. Once a calibration has been performed for each slide lot, alkaline phosphatase activity in unknown samples can be determined from the rate of change in reflectance measured for each unknown test slide.

Validity of a Calibration
Calibration parameters are automatically assessed by the VITROS DT60/DT60 II Chemistry System against a set of quality parameters resident within the analyzer software. Failure to meet any of the pre-defined quality parameters results in a failed calibration. The calibration report should be used in conjunction with quality control results to determine the validity of a calibration.

Reportable (Dynamic) Range Reportable (Dynamic) Range for ALKP DT


Conventional and SI Units (U/L)

15-1500
For out-of-range samples, refer to "Sample Dilution."

Traceability of the Calibration


I I Values assigned to the VITROS Chemistry Products DT Calibrator Kit for alkaline phosphatase are traceable to the alkaline phosphatase method recommended by the International Federation of Clinical Chemistry (IFCC),8'9 adapted to a centrifugal analyzer at 37C.

Quality Control
Procedure Recommendations
| Handle quality control materials as biohazardous material. Choose control levels that check the clinically relevant range. Analyze quality control materials in the same manner as patient samples, before or during patient sample processing. To verify system performance, analyze control materials: - After calibration. - According to local regulations or at least once each day that the test is being performed. - After specified service procedures are performed. Refer to the operator's manual for your VITROS DT60/DT60II System. If control results fall outside your acceptable range, investigate the cause before deciding whether to report patient results. For general quality control recommendations, refer to Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition10 or other published guidelines. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Pub.

No. C-337

Version 1.0

INSTRUCTIONS FOB USE


Expected Values and Reporting Units

ALKPDT
Alkaline Phosphatase

Quality Control Material Selection


VITROS DT Control I & II are recommended for use with the VITROS DT60/DT60 II Chemistry System. Evaluate the performance of other commercial control fluids for compatibility with this test before using for quality control. Control materials other than VITROS DT Controls I & II may show a difference when compared with other alkaline phosphatase methods if they: - Depart from a true human matrix. - Contain high concentrations of preservatives, stabilizers, or other nonphysiological additives. Enzyme activity might also vary with enzyme source, diluent temperature, and activation time during reconstitution. Do not use control materials stabilized with ethylene glycol.

Quality Control Material Preparation and Storage


Refer to the instructions for use for VITROS Chemistry Products DT Control I & II or to other manufacturer's product literature.

Expected Values and Reporting Units


Reference Interval
This reference interval is the central 95% of results from an internal study of 273 apparently healthy adults from a working population (154 females and 119 males).

Reference Interval for ALKP DT


Conventional and SI Units (U/L) 38-126 Each laboratory should confirm the validity of these intervals for the population it serves.

Reporting Units
The VITROS DT60/DT60 II Chemistry System may be programmed to report ALKP DT results in conventional and SI units.

Reporting Units for ALKP DT Conventional and SI Units


U/L

Limitations of the Procedure


Known Interferences
The VITROS ALKP DT Slide method was screened for interfering substances following NCCLS Protocol EP7." The substances listed in the table, when tested at the concentrations indicated, caused the bias shown.

Known Interfering Substances for ALKP DT


Interferent* Bilirubin Methotrexate Nitrofurantoin
*

Interferent Concentration 20 mg/dL 200 ug/mL 40ug/mL (342 umol/L) (440 umol/L) (168 umol/L)

Alkaline Phosphatase Activity Average Bias Conv./SI Units (U/L) Conv./SI Units (U/L) 120 18.0 130 24.0 120 29.0

It is possible that other interfering substances may be encountered. These results are representative; however, your results may differ somewhat due to test-to-test variation. The degree of interference at concentrations other than those listed might not be predictable.

Other Limitations
Some drugs that have significant light absorbance in the region of 400 nm can cause a spectral interference. Certain drugs and clinical conditions are known to alter alkaline phosphatase activity in vivo. For additional information, refer to one of the published summaries.12 13

Version 1.0

Pub. No. C-337

ALKPDT
Alkaline Phosphatase

INSTRUCTIONS FOR USE


Performance Characteristics

Performance Characteristics
Method Comparison
The plot and table show the results of a comparison of samples analyzed on the VITROS DT60 II Chemistry System with those analyzed using the Modified IFCC comparative method8 9 adapted to a centrifugal analyzer at 37C. Testing followed NCCLS Protocol EP9. r "

Method Comparison for ALKP DT: Serum


Conventional and SI Units . 1600'

1200

800 o a:
400

400

800

1200

1600

Comparative Method: Modified IFCC (U/L)

Method Comparison for ALKP DT: Serum


Conventional and SIUnits (U/L) n DT60 II System vs. comparative method 94 Slope 0.99 Correlation Coefficient 0.994 Range of Sample Activity Intercept 5.78 Sy.x 39.10

36-1348

Precision
Precision was evaluated with quality control materials on the VITROS DT60 II System following NCCLS Protocol EP5.15 The data presented are a representation of test performance and are provided as a guideline. Variables such as sample handling and storage, reagent handling and storage, laboratory environment, and system maintenance can affect reproducibility of test results.

Precision for ALKP DT: Serum


Conventional and SI Units (U/L) System VITROS DT60 II Mean Activity 113 446 Within Day SD* 1.9 6.4 Within Lab SD** Within Lab CV%** 3.9 12.7 3.5 2.9 No. Observ. 88 88 No. Days 22 22

Within Day precision was determined using two runs/day with two replications. Within Lab precision was determined using a single lot of slides and calibrating weekly.

Pub.

No. C-337

Version 1.0

|3 VITROS

INSTRUCTIONS FOR USE


References

ALKP DT
Alkaline Phosphatase

References
1. 2. 3. 4. 5. 6. 7. Tietz NW (ed). Fundamentals of Clinical Chemistry, ed. 3. Philadelphia: WB Saunders; 385-386; 1987. NCCLS. Protection of Laboratory Workers from Instrument Biohazards and Infectious Diseases Transmitted by Blood, Body Fluids and Tissue; Approved Guideline. NCCLS Document M29 (OSBN 1-56238). NCCLS, Wayne, PA 19087; 1997. Doumas BT, et al. Differences Between Values for Plasma and Serum in Tests Performed in the Ektachem 700 XR Analyzer, and Evaluation of "Plasma Separator Tubes (PST)." Clin. Chem. 35:151-153; 1989. Calam RR. Specimen Processing Separator Gels: An Update. J Clin Immunoassay. 11:86-90; 1988. Clinical Laboratory Handbook for Patient Preparation and Specimen Handling. Fascicle VI: Chemistry/Clinical Microscopy. Northfield, IL: College of American Pathologists; 1992. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Venipuncture. NCCLS Document H3. Wayne, PA: NCCLS;

1991.
NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Skin Puncture. NCCLS Document H4. Wayne, PA: NCCLS; 1991. 8. Bretaudiere JP, Vassault A, et al. Criteria for establishing a standardized method for determining alkaline phosphatase activity in human serum. Clin. Chem. 23:2263-2274; 1977. 9. Tietz NW, Rinker AD, Shaw L M. IFCC Methods for the Measurement of Catalytic Concentration of Enzymes, Part 5. IFCC Method for Alkaline Phosphatase.. J Clin. Chem., Clin. Biochem. 21:731-748; 1983. 10. NCCLS. Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition. NCCLS Document C24. Wayne, PA: NCCLS; 1999. 11. NCCLS. Interference Testing in Clinical Chemistry. NCCLS Document EP7. Wayne, PA: NCCLS; 1986. 12. Young DS. Effects of Drugs on Clinical Laboratory Tests, ed. 4. Washington D.C.: AACC Press; 1995. 13. Friedman RB, Young DS. Effects of Disease on Clinical Laboratory Tests. Washington, D.C.: AACC Press; 1990. 14. NCCLS. Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline. NCCLS Document EP9. Wayne, RA: NCCLS; 1995. 15. NCCLS. User Evaluation of Precision Performance with Clinical Chemistry Devices. NCCLS Document EP5. Wayne, PA: NCCLS;

1992.

Glossary of Symbols
The Glossary of Symbols table defines the symbols used on the VITROS Chemistry Products packaging and on the VITROS Chemistry Products Assay Sheets. Glossary of Symbols
Do Not Reuse Use by or Expiration Date (YYYY-MM-DD) Lot Number Manufacturer's Serial Number Catalog Number or Product Code Attention: See Instructions for Use. Manufacturer Authorized Representative Contains Sufficient for "n" Tests For In Vitro Diagnostic Use Store Between Consult Instructions for Use Fragile, Handle with Care. Keep Dry This end up

X X

Store At or Below Store At or Above

Version 1.0

Pub. No. C-337

VITRI

A1KPDT
Alkaline Phosphatase

INSTRUCTIONS FOR USE


Revision History

Revision History
Date of Revision 2003-04-30 Version 1.0 Description of Technical Changes* New format New organization and sections consistent with IVD Directive Specimen Storage and Stability - updated all stability values Sample Dilution - added 7% BSA as a diluent Materials Required But Not Provided - added VITROS DT Pipette Quality Control Material Selection - added the statement regarding ethylene glycol Limitations of the Procedure - removed theophylline, added bilirubin, methotrexate, and nitrofurantoin Method Comparison - updated comparison values and plot Precision - updated all values References - added all

The change bars indicate the position of a technical amendment to the text with respect to the previous version of the document.

When this Instructions For Use is replaced, sign and date below and retain as specified by local regulations or laboratory policies, as appropriate.

Signature

Obsolete Date

C
REP

Ortho-Clinical Diagnostics Mandeville House 62 The Broadway Amersham Buckinghamshire HP7 OHJ England Ortho-Clinical Diagnostics, Inc. 100 Indigo Creek Drive Rochester, NY 14626-5101

Ortho-Clinical Diagnostics
company

VITROS is a trademark of Ortho-Clinical Diagnostics, Inc. Ortho-Clinical Diagnostics, Inc., 2003. All rights reserved. Printed in USA.

Pub. No. C-337

Version 1.0

I Products-

VITRCD5
Chemistry I

INSTRUCTIONS FOR USE


VITROS Chemistry Products ALT DT Slides Intended Use

AITDT
Alanine Aminotransferase

For in vitro diagnostic use only. VITROS ALT DT Slides quantitatively measure alanine aminotransferase (ALT) activity in serum and plasma.

Summary and Explanation of the Test


Alanine aminotransferase is present in high activity in liver, skeletal muscle, heart, and kidney. Serum ALT increases rapidly in liver cell necrosis, hepatitis, hepatic cirrhosis, liver tumors, obstructive jaundice, Reye's syndrome, extensive trauma to skeletal muscle, myositis, myocarditis, and myocardial infarction.1

Principles of the Procedure


The VITROS ALT DT Slide method is performed using the VITROS ALT DT Slide and the VITROS Chemistry Products DT Calibrator Kit on VITROS DT60/DT60 II Chemistry Systems. The VITROS ALT DT Slide is a multilayered, analytical element coated on a polyester support. A drop of patient sample is deposited on the slide and is evenly distributed by the spreading layer to the underlying layers. The spreading layer contains the ALT substrates L-alanine and sodium a-ketoglutarate. Alanine aminotransferase catalyzes the transfer of the amino group of L-alanine to a-ketoglutarate to produce pyruvate and glutamate. Lactate dehydrogenase (LDH) then catalyzes the conversion of pyruvate and NADH to lactate and NAD+. The rate of oxidation of NADH is monitored by reflectance spectrophotometry. The rate of change in reflection density is proportional to enzyme activity.

Reaction Sequence
alanine + a-ketoglutarate pyruvate + NADH + H+
ALT pyridoxal-5-phosphate LDH

pyruvate + glutamate - > lactate + NAD*

Test Type and Conditions


Test Type and Conditions for ALT DT
Test Type Rate VITROS DT60/DT60 II Module DTSC/DTSC II Approximate Incubation Time 5 minutes Temperature 37C (98.6F) Wavelength 340 nm Sample Drop Volume 10 ML

Warnings and Precautions


For in vitro diagnostic use only. Take care when handling materials and samples of human origin. Since no test method can offer complete assurance that infectious agents are absent, consider all clinical specimens, controls, and calibrators potentially infectious. Handle specimens, solid and liquid waste, and test components in accordance with local regulations and NCCLS Guideline M292 or other published biohazard safety guidelines. For specific warnings and precautions for calibrators, quality control materials, and other components, refer to the instructions for use for the appropriate VITROS product, or to other manufacturer's product literature.

Version 1.0

Pub. No. C-336

AITDT
Alanine Aminotransferase

INSTRUCTIONS FOR USE


Reagents

Reagents
Slide Diagram

Slide Ingredients
Reactive ingredients per cm
2

1 . 2

Lactate dehydrogenase (porcine muscle, E.C.1.1.1.27) 0.12 U; L-alanine 0.86 mg; sodium a-ketoglutarate 54 ng; nicotinamide adenine dinucleotide, reduced 35 ng; and sodium pyridoxal-5-phosphate 11 ^g.

Other ingredients
Pigment, binders, buffer, surfactants, cross-linking agent and stabilizer.
5

1. Jpper slide mount 2. Spreading layer (BaSO4) sodium a-ketoglutarate L-alanine 3. leagent layer buffer, pH 8.0 > lactate dehydrogenase > NADH pyridoxal-5-phosphate 4. Support layer 6. .ower slide mount

Slide Handling
CAUTION: Do not use slides with damaged or incompletely sealed packaging.

Slide Preparation
IMPORTANT: The slide must reach room temperature, 18 -28 C (64 ~82 F), before the wrapper is opened. Do not use unopened slides that have been at room temperature, 18-28QC (64 -82 F) for >48 hours.

1. 2.

Remove the unopened slide from the box. Warm the unopened slide for 15 minutes. Unopened slides that remain in the sealed wrapper may be returned to refrigerator or freezer storage. 3. Remove the packaging and place the slide into the loading station within 15 minutes after opening.

Slide Storage and Stability


VITROS ALT DT Slides are stable until the expiration date on the carton when they are stored and handled as specified.

Slide Storage and Stability for ALT DT


Slides Unopened

Opened

Storage Condition Room temperature 18-28C (64-82F) Refrigerated 2-8C (36-46F) Frozen <-18C (<0F) Room temperature 18-28C (64-82 F)

Stability <48 hours Until expiration date Until expiration date <15 minutes

Specimen Requirements
WARNING: Handle specimens as biohazardous material.

Specimens Recommended
Serum Plasma: EDTA Heparin Certain collection devices have been reported to affect other analytes and tests3 Confirm that your collection devices are compatible with this test.

IMPORTANT:

Specimens Not Recommended


Do not use hemolyzed specimens.4

Serum and Plasma


Specimen Collection and Preparation
Collect specimens using standard laboratory procedures.56 Patient Preparation No special patient preparation is necessary.

Pub. No. C-336

Version 1.0

INSTRUCTIONS FOR USE


Testing Procedure Special Precautions Centrifuge specimens and remove the serum or plasma from the cellular material within 3 days of collection.7 Specimen Handling and V Storage

ALTDT
Alanine Aminotransferase

Handle specimens as biohazardous material. Handle and store specimens in stoppered containers to avoid contamination and evaporation. Mix samples by gentle inversion and bring to room temperature, 18-28C (64-82F), prior to analysis. Do not freeze the specimen.

IMPORTANT:

Specimen Storage and Stability for ALT DT: Serum and Plasma7
Storage Room temperature Refrigerated Frozen Temperature 18-28C (64O-82F) 2-8C (36-46F) <-18C(<0F) Stability <3 days <1 week Not recommended

Testing Procedure
Materials Provided
VITROS Chemistry Products ALT DT Slides

Materials Required But Not Provided


| . VITROS Chemistry Products DT Calibrator Kit Quality control materials, such as VITROS Chemistry Products DT Control I & II VITROS Chemistry Products 7% BSA or isotonic saline VITROS DT Pipette

Operating Instructions
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System. Bring all fluids and samples to room temperature, 18-28C (64-82F), prior to analysis. IMPORTANT:

Sample Dilution
| If alanine aminotransferase activities exceed the system's reportable (dynamic) range: 1. Dilute the sample with VITROS 7% BSA or isotonic saline. 2. Reanalyze. 3. Multiply the results by the dilution factor to obtain an estimate of the original sample's alanine aminotransferase activity.

Calibration
Required Calibrators
VITROS Chemistry Products DT Calibrator Kit, bottles 1, 2, and 4

Calibrator Preparation, Handling, and Storage


Refer to the instructions for use for VITROS DT Calibrator Kit.

Calibration Procedure
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

When to Calibrate
Calibrate: When the slide lot number changes. When critical system parts are replaced due to service or maintenance. When government regulations require. - For example, in the USA, CLIA regulations require calibration or calibration verification at least once every six months.

Version 1.0

Pub. No. C-336

ALTDT
Alanine Aminotransferase

INSTRUCTIONS FOR USE


Quality Control
The VITROS ALT DT test may also need to be calibrated: If quality control results are consistently outside acceptable range. After certain service procedures have been performed. For additional information, refer to the operator's manual for your VITROS DT60/DT60II Chemistry System.

Calculations
Based on sequential readings of the slide's reflectance at 340 nm over the defined incubation period, a rate of change in reflectance is determined. This rate is used in the software-resident multi-point rate calibration model to compute enzyme activity. Once a calibration has been performed for each slide lot, alanine aminotransferase activity in unknown samples can be determined from the rate of change in reflectance measured for each unknown test slide.

Validity of a Calibration
Calibration parameters are automatically assessed by the VITROS DT60/DT60 II Chemistry System against a set of quality parameters resident within the analyzer software. Failure to meet any of the pre-defined quality parameters results in a failed calibration. The calibration report should be used in conjunction with quality control results to determine the validity of a calibration.

Reportable (Dynamic) Range Reportable (Dynamic) Range for ALT DT


Conventional and SI Units (U/L)

3-950
For out-of-range samples, refer to "Sample Dilution."

Traceability of the Calibration

I
|

Values assigned to the VITROS Chemistry Products DT Calibrator Kit for alanine aminotransferase are traceable to the alanine aminotransferase method recommended by the International Federation of Clinical Chemistry (IFCC),8 adapted to a centrifugal analyzer at 37C.

Quality Control
Procedure Recommendations
WARMING: Handle quality control materials as biohazardous material. Choose control levels that check the clinically relevant range. Analyze quality control materials in the same manner as patient samples, before or during patient sample processing. To verify system performance, analyze control materials: - After calibration. - According to local regulations or at least once each day that the test is being performed. - After specified service procedures are performed. Refer to the operator's manual for your VITROS DT60/DT60 II System. If control results fall outside your acceptable range, investigate the cause before deciding whether to report patient results. For general quality control recommendations, refer to Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition9 or other published guidelines. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Quality Control Material Selection


I | IMPORTANT: VITROS DT Control I & II are recommended for use with the VITROS DT60/DT60II Chemistry System. Evaluate the performance of other commercial control fluids for compatibility with this test before using for quality control.

Control materials other than VITROS DT Controls I & II may show a difference when compared with other alanine aminotransferase methods if they: - Depart from a true human matrix. - Contain high concentrations of preservatives, stabilizers, or other nonphysiological additives. Enzyme activity might also vary with enzyme source, diluent temperature, and activation time during reconstitution. Do not use control materials stabilized with ethylene glycol.

Quality Control Material Preparation and Storage


Refer to the instructions for use for VITROS Chemistry Products DT Control I & II or to other manufacturer's product literature.

Pub. No. C-336

Version 1.0

INSTRUCTIONS FOR USE


Expected Values and Reporting Units

ALTDT
Alanine Aminotransferase

Expected Values and Reporting Units


Reference Interval
These reference intervals are the central 95% of results from an internal study of 2444 apparently healthy adults (547 females and 1897 males).

Reference Interval for ALT DT


Adult Females Males Conventional and SI Units (U/L) . 13-69 . 9-52 21-72

Each laboratory should confirm the validity of these intervals for the population it serves.

Reporting Units
The VITROS DT60/DT60 II Chemistry System may be programmed to report alanine aminotransferase results in units.

Reporting Units for ALT DT


Conventional and SI Units U/L

Limitations of the Procedure


Known Interferences
None identified.

Other Limitations
Certain drugs and clinical conditions are known to alter alanine aminotransferase activity in vivo. For additional information, refer to one of the published summaries.1011

Performance Characteristics
Method Comparison

The plot and table show the results of a comparison of samples analyzed on the VITROS DT60 II System with those analyzed

using the VITROS 950 System. Testing followed NCCLS Protocol EP9.'2

Method Comparison for ALT DT: Serum


10110 Conventional and SI Units
y = x

800
600

401)

200

200

400

600

800

1000

Comparative Method: VITROS 950 System (U/L)

Version 1.0

Pub. No. C-336

A1TDT
Alanine Aminotransferase Method Comparison for ALT DT: Serum

INSTRUCTIONS FOR USE


References

Conventional and SI Units (U/L) n DT60 II System vs. comparative method 59 Correlation Range of Slope Coefficient Sample Activity Intercept 1.03 0.999 6-863 -3.92 Sy.x 11.22

Precision
Precision was evaluated with quality control materials on the VITROS DT60II System following NCCLS Protocol EP5.13 The data presented are a representation of test performance and are provided as a guideline. Variables such as sample handling and storage, reagent handling and storage, laboratory environment, and system maintenance can affect reproducibility of test results.

Precision for ALT DT: Serum


Conventional and SI Units (U/L) System VITROS DT60 II Mean Activity 35 190 Within Day SD* 1.6 1.7 Within Lab SD** 2.0 3.4 Within Lab CV%** 5.7 1.8 No. Observ. 84 84 No. Days 21 21

Within Day precision was determined using two runs/day with two replications. Within Lab precision was determined using a single lot of slides and calibrating weekly.

References
1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. Tietz NW (ed). Fundamentals of Clinical Chemistry, ed. 3. Philadelphia: WB Saunders; 369-371; 1987. NCCLS. Protection of Laboratory Workers from Instrument Biohazards and Infectious Diseases Transmitted by Blood, Body Fluids and Tissue; Approved Guideline. NCCLS Document M29 (OSBN 1-56238). NCCLS, Wayne, PA 19087; 1997. Calam RR. Specimen Processing Separator Gels: An Update. J Clin Immunoassay. 11:86-90; 1988. Young DS. Effects of Preanalytical Variables on Clinical Laboratory Tests. Washington D.C.: AACC Press; 3-7; 1993. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Venipuncture. NCCLS Document H3. Wayne, PA: NCCLS; 1991. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Skin Puncture. NCCLS Document H4. Wayne, PA: NCCLS; 1991. Clinical Laboratory Handbook for Patient Preparation and Specimen Handling. Fascicle VI: Chemistry/Clinical Microscopy. Northfield, IL: College of American Pathologists; 1992. Bergmeyer HU, Horder M, Rej R. Approved Recommendation (1985) on IFCC Methods for the Measurement of Catalytic Concentration of Enzymes. Part 3. IFCC Method for Alanine Aminotransferase. J. Clin. Chem. Clin. Biochem. 24:481; 1986. NCCLS. Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition. NCCLS Document C24. Wayne, PA: NCCLS; 1999. Young DS. Effects of Drugs on Clinical Laboratory Tests, ed. 4. Washington D C : AACC Press; 1995. Friedman RB, Young DS. Effects of Disease on Clinical Laboratory Tests. Washington, D.C.: AACC Press; 1990. NCCLS. Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline. NCCLS Document EP9. Wayne, PA: NCCLS; 1995. NCCLS. User Evaluation of Precision Performance with Clinical Chemistry Devices. NCCLS Document EP5. Wayne, PA: NCCLS; 1992.

Pub. No. C-336

Version 1.0

INSTRUCTIONS FOR USE


Glossary of Symbols

ALTDT
Alanine Aminotransferase

Glossary of Symbols
The Glossary of Symbols table defines the symbols used on the VITROS Chemistry Products packaging and on the VITROS Chemistry Products Assay Sheets.

Glossary of Symbols
Do Not Reuse Use by or Expiration Date (YYYY-MM-DD) Lot Number ^ J | ec | HEP | Manufacturer Authorized Representative Contains Sufficient for "n" Tests For In Vitro Diagnostic Use Store At or Below Store Between Consult Instructions for Use

Fragile, Handle with Care. Keep Dry

SN
REF|

Manufacturer's Serial Number Catalog Number or Product Code Attention: See Instructions for Use.

If

This end up

Store At or Above

Version 1.0

Pub.

No. C-336

VITRCpB

ALTDT
Alanine Aminotransferase

INSTRUCTIONS FOR USE


Revision History

Revision History
Date of Revision 2003-10-01 Version 1.0 Description of Technical Changes* New format New organization and sections consistent with IVD Directive Materials Required But Not Provided and Sample Dilution - added VITROS 7% BSA Quality Control Material Selection - added statements regarding enzyme activity and ethyline glycol Reference Interval - updated all data Known Interferences - removed statement regarding high total protein Method Comparison - updated the comparison and plot Precision - updated all values References - added all except 8

The change bars indicate the position of a technical amendment to the text with respect to the previous version of the document.

When this Instructions For Use is replaced, sign and date below and retain as specified by local regulations or laboratory policies, as appropriate.

Signature

Obsolete Date

C
Ortho-Clinical Diagnostics Mandeville House 62 The Broadway Amersham Buckinghamshire HP7 OHJ England Ortho-Clinical Diagnostics, Inc. 100 Indigo Creek Drive Rochester, NY 14626-5101

Ortho-Clinical Diagnostics
efcMtsH company

VITROS is a trademark of Ortho-Clinical Diagnostics, Inc. Ortho-Clinical Diagnostics, Inc., 2003. All rights reserved. Printed in USA.

Pub. No. C-336

Version 1.0

(Products

VITRCD5
Chemistry I

INSTRUCTIONS FOR USE


VITROS Chemistry Products AMYL DT Slides Intended Use
For in vitro diagnostic use only. VITROS AMYL DT Slides quantitatively measure amylase (AMYL) activity in serum and plasma.

AMY1 DT
Amylase

Summary and Explanation of the Test


Amylase is an amylolytic digestive enzyme produced by the exocrine pancreas and salivary glands. Amylase is increased in acute pancreatitis, pancreatic abscess or pseudocyst, pancreatic trauma, amyloidosis, pancreatic neoplasm, common-bile-duct obstruction, and after thoracic surgery. Increased amylase activity may be found in mumps parotitis and renal insufficiency.1

Principles of the Procedure


The VITROS AMYL DT Slide method is performed using the VITROS AMYL DT Slide and the VITROS Chemistry Products DT Calibrator Kit on VITROS DT60/DT60 II Chemistry Systems. The VITROS AMYL DT Slide is a multilayered, analytical element coated on a polyester support. A drop of patient sample is deposited on the slide and is evenly distributed by the spreading layer to the underlying layers. The spreading layer contains the dyed starch substrate (dye covalently linked to amylopectin) for the reaction. The amylase in the sample catalyzes the hydrolysis of this dyed starch into smaller dyed saccharides. These dyed saccharides diffuse into the underlying reagent layer. The reflection density is proportional to the activity of amylase present in the sample.

Reaction Sequence
dyed amylopectin
amylase

->

dyed saccharides

Test Type and Conditions


Test Type and Conditions for AMYL DT
Test Type Endpoint VITROS DT60/DT60 II Module DT60/DT60 II Approximate Incubation Time 5 minutes Temperature 37C (98.6F) Wavelength 555 nm Sample Drop Volume 10 uL

Warnings and Precautions


For in vitro diagnostic use only. Take care when handling materials and samples of human origin. Since no test method can offer complete assurance that infectious agents are absent, consider all clinical specimens, controls, and calibrators potentially infectious. Handle specimens, solid and liquid waste, and test components in accordance with local regulations and NCCLS Guideline M292 or other published biohazard safety guidelines. For specific warnings and precautions for calibrators, quality control materials, and other components, refer to the instructions for use for the appropriate VITROS product, or to other manufacturer's product literature.

Version 1.0

Pub. No. C-311

AMYLDT
Amylase

INSTRUCTIONS FOR USE


Reagents

Reagents
Slide Diagram

Slide .Ingredients

Reactive ingredients per cm


Dyed amylopectin 320 ug.

Other ingredients
Pigment, binders, buffers, mordant, surfactants and stabilizer.

1. Upper slide mount 2. Spreading layer (BaSO,>) dyed amylopectin buffer, pH 7.2 3. Reagent layer buffer, pH 7.2 4. Support layer 5. Lower slide mount

Slide Handling
Do not use slides with damaged or incompletely sealed packaging.

Slide Preparation
IMPORTANT: The slide must reach room temperature, 18-28 C (64 "-82 F), before the wrapper is opened. Do not use unopened slides that have been at room temperature, 18-28C (64-82F) for >48 hours. 1. 2. Remove the unopened slide from the box. Warm the unopened slide for 15 minutes. Unopened slides that remain in the sealed wrapper may be returned to refrigerator or freezer storage. 3. Remove the packaging and place the slide into the loading station within 15 minutes after opening.

Slide Storage and Stability


VITROS AMYL DT Slides are stable until the expiration date on the carton when they are stored and handled as specified.

Slide Storage and Stability for AMYL DT


Slides Unopened Storage Condition Room temperature 18-28C (64-82F) Refrigerated 2-8C (36-46F) Frozen <-18C(<0F) Room temperature 18-28C (64-82F) Stability <48 hours Until expiration date Until expiration date <15 minutes

Opened

Specimen Requirements
WARNING: Handle specimens as biohazardous material.

Specimens Recommended
Serum Plasma: Heparin3 Plasma activities are approximately 20 U/L higher than serum activities.3 Certain collection devices have been reported to affect other analytes and tests.4 Confirm that your collection devices are compatible with this test

iTOTE;

Specimens Not Recommended


Plasma:5 Citrate EDTA Fluoride oxalate

Pub. No. C-311

Version 1.0

INSTRUCTIONS FOR USE


Testing Procedure Serum and Plasma Specimen Collection and Preparation
Collect specimens using standard laboratory procedures.6 7 Patient Preparation No special patient preparation is necessary. Special Precautions Centrifuge specimens and remove the serum from the cellular material within 4 hours of collection.8 Storage

AMY1DT
Amylase

Specimen Handling and

Handle specimens as biohazardous material. Handle and store specimens in stoppered containers to avoid contamination and evaporation. Mix samples by gentle inversion and bring to room temperature, 18-28C (64-82F), prior to analysis. Do not freeze the specimen.

IMPORTANT;

Specimen Storage and Stability for AMYL DT: Serum and Plasma8 Storage Temperature Stability
Room temperature Refrigerated Frozen 18O-28C(64O-82F) 2-8C (36-46F) <-18C(<0F) <7 days <1 month Not recommended

Testing Procedure
Materials Provided
VITROS Chemistry Products AMYL DT Slides

Materials Required But Not Provided


VITROS Chemistry Products DT Calibrator Kit Quality control materials, such as VITROS Chemistry Products DT Control I & II Isotonic saline VITROS DT Pipette

Operating Instructions
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System. Bring all fluids and samples to room temperature, 18-28C (64-82F), prior to analysis. IMPORTANT:

Sample Dilution
I If amylase activities exceed the system's reportable (dynamic) range: 1. Dilute the sample with a patient sample with low amylase activity or with isotonic saline. 2. Reanalyze. 3. If necessary, correct for amylase activity in the diluent. 4. Multiply the results by the dilution factor to obtain an estimate of the original sample's amylase activity.

Calibration
Required Calibrators
VITROS Chemistry Products DT Calibrator Kit, bottles 1, 2, and 3

Calibrator Preparation, Handling, and Storage


Refer to the instructions for use for VITROS DT Calibrator Kit.

Version 1.0

Pub. No. C-311

AMVLDT
Amylase Calibration Procedure

INSTRUCTIONS FOR USE


Quality Control

Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

When to Calibrate
Calibrate: When the slide lot number changes. When critical system parts are replaced due to service or maintenance. When government regulations require. - For example, in the USA, CLIA regulations require calibration or calibration verification at least once every six months. The VITROS AMYL DT test may also need to be calibrated: If quality control results are consistently outside acceptable range. After certain service procedures have been performed. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Calculations
Reflectance from the slide is measured at 555 nm after the fixed incubation time. Once a calibration has been performed for each slide lot, amylase activity in unknown samples can be determined using the software-resident endpoint colorimetric math model and the response obtained from each unknown test slide.

Validity of a Calibration
Calibration parameters are automatically assessed by the VITROS DT60/DT60 II Chemistry System against a set of quality parameters resident within the analyzer software. Failure to meet any of the pre-defined quality parameters results in a failed calibration. The calibration report should be used in conjunction with quality control results to determine the validity of a calibration.

Re portable (Dynamic) Range Reportable (Dynamic) Range for AMYL DT


Conventional and SI Units (U/L)

5-900
For out-of-range samples, refer to "Sample Dilution."

Traceability of the Calibration


I | Values assigned to the VITROS Chemistry Products DT Calibrator Kit for amylase are traceable to the paranitrophenol maltopentaoside method9 at 37C.

Quality Control
Procedure Recommendations
| WARN i NO; Handle quality control materials as biohazardous material. Choose control levels that check the clinically relevant range. Analyze quality control materials in the same manner as patient samples, before or during patient sample processing. To verify system performance, analyze control materials: - After calibration. - According to local regulations or at least once each day that the test is being performed. - After specified service procedures are performed. Refer to the operator's manual for your VITROS DT60/DT60 II System. If control results fall outside your acceptable range, investigate the cause before deciding whether to report patient results. For general quality control recommendations, refer to Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition10 or other published guidelines. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Pub. No. C-311

Version 1.0

[3 VITROS

INSTRUCTIONS FOR USE


Expected Values and Reporting Units Quality Control Material Selection
I I IMPORTANT; VITROS DT Control I & II are recommended for use with the VITROS DT60/DT60 II Chemistry System. Evaluate the performance of other commercial control fluids for compatibility with this test before using for quality control.

AMYLDT
Amylase

Control materials other than VITROS DT Controls I & II may show a difference when compared with other Amylase methods if they: - Depart from a true human matrix. - Contain high concentrations of preservatives, stabilizers, or other nonphysiological additives. Serum controls with porcine or bovine amylase may give lower values that may vary from method to method.11 Enzyme activity might also vary with enzyme source, diluent temperature, and activation time during reconstitution. Do not use control materials stabilized with ethylene glycol.

Quality Control Material Preparation and Storage


Refer to the instructions for use for VITROS Chemistry Products DT Control I & II or to other manufacturer's product literature.

Expected Values and Reporting Units


Reference Interval
The serum reference interval is the central 95% of results from an internal study of 98 apparently healthy individuals from a working population (55 females and 43 males). No significant differences between results from the male and female populations were observed.

Reference Interval for AWIYL DT


Conventional and SI Units (U/L)

30-110
* ** Plasma concentrations are approximately 20 U/L higher than serum concentrations.3 Adults; normal intervals for children <1 year old are lower. ' 2

Each laboratory should confirm the validity of these intervals for the population it serves.

Reporting Units
The VITROS DT60/DT60 II Chemistry System may be programmed to report Amylase results in conventional and SI units.

Reporting Units for AMYL DT

Conventional and SI Units U/L

Limitations of the Procedure


Known Interferences
None identified.

Other Limitations
Certain drugs and clinical conditions are known to alter amylase activity in vivo. For additional information, refer to one of the published summaries. 13 ' u

Version 1.0

Pub. No. C-311

AMY1DT
Amylase

INSTRUCTIONS FOR USE


Performance Characteristics

Performance Characteristics
Method Comparison

The plot and table show the results of a comparison of samples analyzed on the VITROS DT60 II System with those analyzed

using the Paranitrophenol Maltopentaoside comparative method.9

Method Comparison for AMYL DT: Serum


Conventional and SI Units
1000

900 ' 800" 700 600 " 500 400 ' 300 200 100 1000 200 400 600 800 0 0 Comparative Method: Paranitrophenol Maltopentaoside
(U/L)

Method Comparison for AMYL DT: Serum


Conventional and SI Units (U/L) Correlation Range of Slope Coefficient Sample Activity Intercept Sy.x 0.98 0.990 42-888 3.89 35.99

n DT60 II System vs. comparative method 69

Precision
Precision was evaluated with quality control materials on the VITROS DT60 II System following NCCLS Protocol EP5.16 The data presented are a representation of test performance and are provided as a guideline. Variables such as sample handling and storage, reagent handling and storage, laboratory environment, and system maintenance can affect reproducibility of test results.

Precision for AMYL DT: Serum


Conventional and SI Units (U/L) System VITROS DT60 II Mean Activity 100 422 Within Day SD* 4.5 13.3 Within Lab SD** Within Lab CV%** 5.2 16.1 5.2 3.8 No. Observ. 88 88 No. Days 22 22

Within Day precision was determined using two runs/day with two replications. Within Lab precision was determined using a single lot of slides and calibrating weekly.

Pub. No. C-311

Version 1.0

INSTRUCTIONS FOR USE


References

AMYLDT
Amylase

References
1. 2. 3. 4. 5. 6. 7. Tietz NW (ed). Fundamentals of Clinical Chemistry, ed. 3. Philadelphia: WB Saunders; 394-395; 1987. NCCLS. Protection of Laboratory Workers from Instrument Biohazards and Infectious Diseases Transmitted by Blood, Body Fluids and Tissue; Approved Guideline. NCCLS Document M29 (OSBN 1-56238). NCCLS, Wayne, PA 19087; 1997. Doumas BT, et al. Differences Between Values for Plasma and Serum in Tests Performed in the Ektachem 700 XR Analyzer, and Evaluation of "Plasma Separator Tubes (PST)." Clin. Chem. 35:151-153; 1989. Calam RR. Specimen Processing Separator Gels: An Update. J Clin Immunoassay. 11:86-90; 1988. Tietz NW (ed). Fundamentals of Clinical Chemistry, ed. 5. Philadelphia: WB Saunders; 376; 2001. NCCLS. Procedures for the. Collection of Diagnostic Blood Specimens by Venipuncture. NCCLS Document H3. Wayne, PA: NCCLS;

1991.
NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Skin Puncture. NCCLS Document H4. Wayne, PA: NCCLS; 1991. 8. Clinical Laboratory Handbook for Patient Preparation and Specimen Handling. Fascicle VI: Chemistry/Clinical Microscopy. Northfield, IL: College of American Pathologists; 1992. 9. Mauck LA. A Kinetic Colorimetric Method for the Determination of Total Amylase Activity in Serum. Clin. Chem. 31:1007; 1985. 10. NCCLS. Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition. NCCLS Document C24. Wayne, PA: NCCLS; 1999. 11. Lee VW, Willis C. Activity of Human and Nonhuman Amylases on Different Substrates Used in Enzymatic Kinetic Assay Methodsa Pitfall in Interlaboratory Quality Control. Am. J. Clin. Path. 77:290-296; 1982. 12. Gillard BK, Simbala JA, Goodnick L. Reference Intervals for Amylase Isoenzymes in Serum and Plasma of Infants and Children. Clin. Chem. 29:1119; 1983. 13. Young DS. Effects of Drugs on Clinical Laboratory Tests, ed. 4. Washington D C : AACC Press; 1995. 14. Friedman RB, Young DS. Effects of Disease on Clinical Laboratory Tests. Washington, D.C.: AACC Press; 1990. 15. NCCLS. User Evaluation of Precision Performance with Clinical Chemistry Devices. NCCLS Document EP5. Wayne, PA: NCCLS;

1992.

Glossary of Symbols
The Glossary of Symbols table defines the symbols used on the VITROS Chemistry Products packaging and on the VITROS Chemistry Products Assay Sheets.

Glossary of Symbols
Do Not Reuse Use by or Expiration Date (YYYY-MM-DD) Lot Number CM OlM _ Kti" Manufacturer's Serial Number Catalog Number or Product Code Attention: See Instructions for Use. ^m EC I REP I Manufacturer Authorized Representative Contains Sufficient for "n" Tests For In Vitro Diagnostic Use Store At or Below Store Between Consult Instructions for Use Fragile; Handle with Care. Keep Dry This end up

Store At or Above

Version 1.0

Pub. No. C-311

VITRI

AMYLDT
Amylase

INSTRUCTIONS FOR USE


Revision History

Revision History
Date of Revision 2003-08-11 Description of Technical Changes" New format > New organization and sections consistent with IVD Directive > Specimen Storage and Stability - updated stability > Sample Dilution - added isotonic saline as a diluent; removed 2% bovine serum albumin in saline, pH 7.4 Method Comparison - updated the data and plot Precision - updated all data > References - added all but 3 The change bars indicate the position of a technical amendment to the text with respect to the previous version of the document. Version 1.0

When this Instructions For Use is replaced, sign and date below and retain as specified by local regulations or laboratory policies, as appropriate.

Signature

Obsolete Date

C
REP

Ortho-Clinical Diagnostics Mandeville House 62 The Broadway Amersham Buckinghamshire HP7 OHJ England Ortho-Clinical Diagnostics, Inc. 100 Indigo Creek Drive Rochester, NY 14626-5101

' Ortho-Clinical Diagnostics


^ofaMOH company

VITROS is a trademark of Ortho-Clinical Diagnostics, Inc. Ortho-Clinical Diagnostics, Inc., 2003. All rights reserved. Printed in USA.

Pub. No. C-311

Version 1.0

[Products-

ViTRmS
Chemistry!

INSTRUCTIONS FOR USE


VITROS Chemistry Products AST DT Slides Intended Use

ASTDT
Aspartate Aminotransferase

For in vitro diagnostic use only. VITROS AST DT Slides quantitatively measure aspartate aminotransferase (AST) activity in serum and plasma.

Summary and Explanation of the Test


Aspartate aminotransferase is present in high activity in heart, skeletal muscle, and liver. Increased serum AST activity commonly follows myocardial infarction, pulmonary emboli, skeletal muscle trauma, alcoholic cirrhosis, viral hepatitis, and druginduced hepatitis.1

Principles of the Procedure


The VITROS AST DT Slide method is performed using the VITROS AST DT Slide and the VITROS Chemistry Products DT Calibrator Kit on VITROS DT60/DT60 II Chemistry Systems. The VITROS AST DT Slide is a multilayered, analytical element coated on a polyester support. A drop of patient sample is deposited on the slide and is evenly distributed by the spreading layer to the underlying layers. The spreading layer contains the AST substrates aspartate and a-ketoglutarate. In the assay for aspartate aminotransferase, the amino group of L-aspartate is transferred to a-ketoglutarate in the presence of pyridoxal-5-phosphate (P-5-P) to produce glutamate and oxaloacetate. Malate dehydrogenase (MDH) then catalyzes the conversion of oxaloacetate and NADH to malate and NAD*. The rate of oxidation of NADH is monitored by reflectance spectrophotometry. The rate of change in reflection density is proportional to enzyme activity in the sample.

Reaction Sequence
aspartate + a-ketoglutarate oxaloacetate + NADH + H* AST pyridoxal-5-phosphate MDH

->

oxaloacetate + glutamate

- > malate + NAD+

Test Type and Conditions


Test Type and Conditions for AST DT
Test Type Rate VITROS DT60/DT60 II Module DTSC/DTSC II Approximate Incubation Time 5 minutes Temperature 37C (98.6F) Wavelength 340 nm Sample Drop Volume 10 ML

Warnings and Precautions


For in vitro diagnostic use only. Take care when handling materials and samples of human origin. Since no test method can offer complete assurance that infectious agents are absent, consider all clinical specimens, controls, and calibrators potentially infectious. Handle specimens, solid and liquid waste, and test components in accordance with local regulations and NCCLS Guideline M292 or other published biohazard safety guidelines. For specific warnings and precautions for calibrators, quality control materials, and other components, refer to the instructions for use for the appropriate VITROS product, or to other manufacturer's product literature.

Version 1.0

Pub. No. C-338

ASTDT
Aspartate Aminotransferase

INSTRUCTIONS FOR USE


Reagents

Reagents
Slide Diagram

Slide Ingredients
Reactive ingredients per cm
2

Lactate dehydrogenase (porcine muscle, E.C.1.1.1.27) 0.12 U; malate dehydrogenase (porcine heart, E.C. 1.1.1.3) 0.11 U; sodium aspartate 0.5 mg; sodium a-ketoglutarate 32 ug; nicotinamide adenine dinucleotide, reduced 36 ug; and sodium pyridoxal-5-phosphate 16 ug.

. -i-

Other ingredients
Pigment, binders, buffer, surfactants, cross-linking agent and stabilizer.

- s

1. Upper slide mount 2. Spreading layer (BaSOJ sodium u-ketoglutarate L-aspartate 3. 1Reagent layer buffer, pH 8.0 lactate dehydrogenase malate dehydrogenase NADH pyridoxal-5-phosphate 4. Support layer 6. Lower slide mount

Slide Handling
Do not use slides with damaged or incompletely sealed packaging.

Slide Preparation
The slide must reach room temperature, 18 -28 C (64 "-82 f), before the wrapper is opened. Do not use unopened slides that have been at room temperature, 18 -28 C (64-82F) for >48 hours. 1. 2. Remove the unopened slide from the box. Warm the unopened slide for 15 minutes. Unopened slides that remain in the sealed wrapper may be returned to refrigerator or freezer storage. 3. Remove the packaging and place the slide into the loading station within 15 minutes after opening.

Slide Storage and Stability


VlTROS AST DT Slides are stable until the expiration date on the carton when they are stored and handled as specified.

Slide Storage and Stability for AST DT Slides Storage Condition


Unopened Room temperature Refrigerated Frozen Room temperature 18-28C (64-82F) 2-8C (36-46F) <-18C (<0F) 18-28C (64-82F)

Opened

Stability <48 hours Until expiration date Until expiration date <15 minutes

Specimen Requirements
WARMS: Handle specimens as biohazardous material.

Specimens Recommended
Serum Plasma: Heparin Certain collection devices have been reported to affect other analytes and tests.: Confirm that your collection devices are compatible with this test.

I
'

MPORTANT:

Specimens Not Recommended


Plasma: EDTA Citrate Fluoride oxalate Do not use hemolyzed specimens because of high levels of AST activity in erythrocytes.4

Pub. No. C-338

Version 1.0

INSTRUCTIONS FOR USE


Testing Procedure Serum and Plasma

AST OT
Aspartate Aminotransferase

Specimen Collection and Preparation Collect specimens using standard laboratory procedures.5 6 Due to the very low density of platelets, it is important to centrifuge plasma specimens at a minimum of 1000 X g for a minimum of ten minutes in order to avoid contamination of plasma with AST derived from platelets. Patient Preparation No special patient preparation is necessary. Special Precautions Plasma, specimens must be collected in tubes that are at least half full. Smaller volumes may give falsely high AST results.7 Avoid agitation or mixing of plasma samples after centrifugation. Re-suspension of platelets into previously centrifuged plasma may lead to artificially elevated AST results because of high AST activity in platelets.4 Centrifuge specimens and remove the serum or plasma from the cellular material within 3 days of collection.8 Specimen Handling and Storage Handle specimens as biohazardous material. Handle and store specimens in stoppered containers to avoid contamination and evaporation. Mix samples by gentle inversion and bring to room temperature, 18-28C (64-82F), prior to analysis. Do not freeze the specimen.

Specimen Storage and Stability for AST DT: Serum and Plasma8
Storage Room temperature Refrigerated Frozen Temperature 18-28C(64-82F) 2-8C (36-46F) <-18C(<0F) Stability <3 days <7 days <3 months

Testing Procedure
Materials Provided
VITROS Chemistry Products AST DT Slides

Materials Required But Not Provided


. VITROS Chemistry Products DT Calibrator Kit Quality control materials, such as VITROS Chemistry Products DT Control I & II VITROS Chemistry Products 7% BSA or isotonic saline VITROS DT Pipette

Operating Instructions
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System. Bring all fluids and samples to room temperature, 18-28C (64-82F), prior to analysis.

I
|

IMPORTANT:

Sample Dilution
If aspartate aminotransferase activities exceed the system's reportable (dynamic) range: 1. Dilute the sample with VITROS 7% BSA or isotonic saline. 2. Reanalyze. 3. Multiply the results by the dilution factor to obtain an estimate of the original sample's aspartate aminotransferase activity.

Calibration
Required Calibrators
VITROS Chemistry Products DT Calibrator Kit, bottles 1, 2, and 4

Calibrator Preparation, Handling, and Storage


Refer to the instructions for use for VITROS DT Calibrator Kit.

Version 1.0

Pub. No. C-338

AST DT
Aspartate Aminotransferase Calibration Procedure

INSTRUCTIONS FOR USE


Quality Control
Refer to the operator's manual for your VlTROS DT60/DT60 II Chemistry System.

When to Calibrate
Calibrate: When the slide lot number changes. When critical system parts are replaced due to service or maintenance. When government regulations require. - For example, in the USA, CLIA regulations require calibration or calibration verification at least once every six months. The VlTROS AST DT test may also need to be calibrated: If quality control results are consistently outside acceptable range. After certain service procedures have been performed. For additional information, refer to the operator's manual for your VlTROS DT60/DT60 II Chemistry System.

Calculations
Based on sequential readings of the slide's reflectance at 340 nm over the defined incubation period, a rate of change in reflectance is determined. This rate is used in the software-resident multi-point rate calibration model to compute enzyme activity. Once a calibration has been performed for each slide lot, aspartate aminotransferase activity in unknown samples can be determined from the rate of change in reflectance measured for each unknown test slide.

Validity of a Calibration
Calibration parameters are automatically assessed by the VlTROS DT60/DT60 II Chemistry System against a set of quality parameters resident within the analyzer software. Failure to meet any of the pre-defined quality parameters results in a failed calibration. The calibration report should be used in conjunction with quality control results to determine the validity of a calibration.

Reportable (Dynamic) Range Reportable (Dynamic) Range for AST DT


Conventional and SI Units (U/L)

4-950
For out-of-range samples, refer to "Sample Dilution."

Traceability of the Calibration


I I Values assigned to the VlTROS Chemistry Products DT Calibrator Kit for aspartate aminotransferase are traceable to the aspartate aminotransferase method recommended by the International Federation of Clinical Chemistry (IFCC),9 adapted to a centrifugal analyzer at 37C.

Quality Control
Procedure Recommendations
| WARNING: Handle quality control materials as biohazardous material. Choose control levels that check the clinically relevant range. Analyze quality control materials in the same manner as patient samples, before or during patient sample processing. To verify system performance, analyze control materials: - After calibration. - According to local regulations or at least once each day that the test is being performed. - After specified service procedures are performed. Refer to the operator's manual for your VlTROS DT60/DT60II System. If control results fall outside your acceptable range, investigate the cause before deciding whether to report patient results. For general quality control recommendations, refer to Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition or other published guidelines. For additional information, refer to the operator's manual for your VlTROS DT60/DT60 II Chemistry System.

Pub. No. C-338

Version 1.0

INSTRUCTIONS FOR USE


Expected Values and Reporting Units

ASTDT
Aspartate Aminotransferase

Quality Control Material Selection

I
I

IMPORTANT;

VITROS DT Control I & II are recommended for use with the VITROS DT60/DT60 II

Chemistry System. Evaluate the performance of other commercial control fluids for compatibility with this test before using for quality control. Control materials other than VITROS DT Controls I & II may show a difference when compared with other aspartate aminotransferase methods if they: - Depart from a true human matrix. - Contain high concentrations of preservatives, stabilizers, or other nonphysiological additives. Enzyme activity might also vary with enzyme source, diluent temperature, and activation time during reconstitution. Do not use control materials stabilized with ethylene glycol.

Quality Control Material Preparation and Storage


Refer to the instructions for use for VITROS Chemistry Products DT Control I & II or to other manufacturer's product literature.

Expected Values and Reporting Units


Reference Interval
These reference intervals are the central 95% of results from an internal study of 189 apparently healthy adults from a working population (90 females and 99 males). Reference Interval for AST DT Conventional and SI Units (U/L) Adult Females Males 15-46 14-36 17-59

Each laboratory should confirm the validity of these intervals for the population it serves.

Reporting Units
The VITROS DT60/DT60 II Chemistry System may be programmed to report aspartate aminotransferase results in conventional and SI units.

Reporting Units for AST DT


Conventional and SI Units U/L

Limitations of the Procedure


Known Interferences

Heparin at activities greater than 50 U/mL cause AST results to be falsely high.7 Refer to "Specimen Collection and Preparation."

Other Limitations
Certain drugs and clinical conditions are known to alter aspartate aminotransferase activity in vivo. For additional information, refer to one of the published summaries.1112

Version 1.0

Pub. No. C-338

VITRCpSgJ

ASTDT
Aspartate Aminotransferase

INSTRUCTIONS FOR USE


Performance Characteristics

Performance Characteristics
Method Comparison

The plot and table show the results of a comparison of samples analyzed on the VlTROS DT60 II System with those analyzed using the VlTROS 950 System. Testing followed NCCLS Protocol EP9.13 Method Comparison for A S T DT: Serum Conventional and SI Units 1000 '

<! =

800 600
400

I
o a. >

200

200

400

600

800 1000

Comparative Method: VlTROS 950 System (U/L)

Method Comparison for AST DT: Serum


Conventional and SI Units (U/L) n DT60 II System vs. 950 System 73 Slope 1.01 Correlation Coefficient 0.997 Range of Sample Activity Intercept -8.46 Sy.x 21.68

15-848

Precision
Precision was evaluated with quality control materials on the VlTROS DT60 II System following NCCLS Protocol EP5.14 The data presented are a representation of test performance and are provided as a guideline. Variables such as sample handling and storage, reagent handling and storage, laboratory environment, and system maintenance can affect reproducibility of test results.

Precision for AST DT: Serum


Conventional and SIUnits (U/L) System VlTROS DT60 II Mean Activity 40 193 Within Day SD * 1.7 3.2 Within Lab SD** Within Lab CV%** 3.0 4.1 7.4 2.1 No. Observ. 84 84 No. Days 21
21

Within Day precision was determined using two runs/day with two replications. Within Lab precision was determined using a single lot of slides and calibrating weekly.

Pub. No. C-338

Version 1.0

INSTRUCTIONS FOR USE


References

ASTDT
Aspartate Aminotransferase

References
1. 2. 3. 4. 5. 6. 7. Tietz NW(ed). Fundamentals of Clinical Chemistry, ed. 3. Philadelphia: WB Saunders; 369-371; 1987. NCCLS. Protection of Laboratory Workers from Instrument Biohazards and Infectious Diseases Transmitted by Blood, Body Fluids and Tissue; Approved Guideline. NCCLS Document M29 (OSBN 1-56238). NCCLS, Wayne, PA 19087; 1997. Calam RR. Specimen Processing Separator Gels: An Update. J Clin Immunoassay. 11:86-90; 1988. Young DS. Effects of Preanalytical Variables on Clinical Laboratory Tests, ed. 2. Washington D C : AACC Press; 3-69, 3-70; 1997. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Venipuncture. NCCLS Document H3. Wayne, PA: NCCLS;

1991.
NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Skin Puncture. NCCLS Document H4. Wayne, PA: NCCLS;

1991.
Berg JD, Romano G, Bayley NF, Buckley BWI. Heparin Interferes with Aspartate Aminotransferase Activity Determined in the Ektachem 700. Clin Chem. 34:174; 1988. 8. Clinical Laboratory Handbook for Patient Preparation and Specimen Handling. Fascicle VI: Chemistry/Clinical Microscopy. Northfield, IL: College of American Pathologists; 1992. 9. Bergmeyer H U, Horder M, Rej R. Approved Recommendation on IFCC Methods for the Measurement of Catalytic Concentration of Enzymes. Part 2, IFCC Method for Aspartate Aminotransferase. J. Clin. Chem. Clin. Biochem. 24:497-510; 1986. 10. NCCLS. Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition. NCCLS Document C24. Wayne, PA: NCCLS; 1999. 11. Young DS. Effects of Drugs on Clinical Laboratory Tests, ed. 4. Washington D.C.: AACC Press; 1995. 12. Friedman RB, Young DS. Effects of Disease on Clinical Laboratory Tests. Washington, D C : AACC Press; 1990. 13. NCCLS. Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline. NCCLS Document EP9. Wayne, PA:

NCCLS; 1995.
14. NCCLS. User Evaluation of Precision Performance with Clinical Chemistry Devices. NCCLS Document EP5. Wayne, PA: NCCLS;

1992.

Glossary of Symbols
The Glossary of Symbols table defines the symbols used on the VITROS Chemistry Products packaging and on the VITROS Chemistry Products Assay Sheets.

Glossary of Symbols
Do Not Reuse Use by or Expiration Date (YYYY-MM-DD) Lot Number Manufacturer Authorized Representative Contains Sufficient for "n" Tests For In Vitro Diagnostic Use Store At or Below

Store Between
Consult Instructions for Use Fragile, Handle with Care. Keep Dry This end up

SN
"RIFI

Manufacturer's Serial Number Catalog Number or Product Code Attention: See Instructions for Use.

Store At or Above

Version 1.0

Pub. No. C-338

ASTDT
Aspartate Aminotransferase

INSTRUCTIONS FOR USE


Revision History

Revision History
Date of Revision 2003-10-01 Version 1.0 Description of Technical Changes* New format New organization and sections consistent with IVD Directive Specimens Not Recommended - added citrate Materials Required But Not Provided and Sample Dilution - added VITROS 7% BSA Quality Control Material Selection - added statements regarding enzyme activity and ethylene glycol Reference Interval - updated all data Known Interferences - added statement regarding heparin; removed statement regarding high total protein Method Comparison - updated the comparison and plot Precision - upated all values References - added all

The change bars indicate the position of a technical amendment to the text with respect to the previous version of the document.

When this Instructions For Use is replaced, sign and date below and retain as specified by local regulations or laboratory policies, as appropriate.

Signature

Obsolete Date

C
EC

Ortho-Clinical Diagnostics Mandeville House 62 The Broadway Amersham Buckinghamshire HP7 OHJ England Ortho-Clinical Diagnostics, Inc. 100 Indigo Creek Drive Rochester, NY 14626-5101

'Ortho-Clinical Diagnostics
M|jefcMtBn company

VITROS is a trademark of Ortho-Clinical Diagnostics, Inc. Ortho-Clinical Diagnostics, Inc., 2003. All rights reserved. Printed in USA.

Pub. No. C-338

Version 1.0

VITRCDS
Chemistry!

INSTRUCTIONS FOR USE


VITROS Chemistry Products BUN/UREA DT Slides Intended Use

RUN/ UREADT
Urea Nitrogen

For in vitro diagnostic use only. VITROS BUN/UREA DT Slides quantitatively measure urea concentration, reported either as urea nitrogen (BUN) or as urea (UREA), in serum and plasma.

Summary and Explanation of the Test


The major pathway of nitrogen excretion is in the form of urea that is synthesized in the liver, released into the blood, and cleared by the kidneys. A high serum urea nitrogen occurs in glomerulonephritis, shock, urinary tract obstruction, pyelonephritis, and other causes of acute and chronic renal failure. Severe congestive heart failure, hyperalimentation, diabetic ketoacidosis, dehydration, and bleeding from the gastrointestinal tract elevate urea nitrogen. Low urea nitrogen often occurs in normal pregnancy, with decreased protein intake, in acute liver failure, and with intravenous fluid administration.1

Principles of the Procedure


The VITROS BUN/UREA DT Slide method is performed using the VITROS BUN/UREA DT Slide and the VITROS Chemistry Products DT Calibrator Kit on VITROS DT60/DT60 II Chemistry Systems. The VITROS BUN/UREA DT Slide is a multilayered, analytical element coated on a polyester support. A drop of patient sample is deposited on the slide and is evenly distributed by the spreading layer to the underlying layers. Water and nonproteinaceous components then travel to the underlying reagent layer, where the urease reaction generates ammonia. The semipermeable membrane allows only ammonia to pass through to the color-forming layer, where it reacts with the indicator to form a dye. The reflection density of the dye is measured and is proportional to the concentration of urea in the sample.

Reaction Sequence
H2NCONH2 + H2O NH3 + ammonia indicator - > 2NH3 + CO2

-> dye

Test Type and Conditions


Test Type and Conditions for BUN/UREA DT
Test Type Colorimetric VITROS DT60/DT60 II Module .DT60/DT60II Approximate Incubation Time 5 minutes Temperature 37C (98.6F) Wavelength 660 nm Sample Drop Volume 10 ML

Warnings and Precautions


For in vitro diagnostic use only. Take care when handling materials and samples of human origin. Since no test method can offer complete assurance that infectious agents are absent, consider all clinical specimens, controls, and calibrators potentially infectious. Handle specimens, solid and liquid waste, and test components in accordance with local regulations and NCCLS Guideline M292 or other published biohazard safety guidelines. For specific warnings and precautions for calibrators, quality control materials, and other components, refer to the instructions for use for the appropriate VITROS product, or to other manufacturer's product literature.

Version 1.0

Pub.

No. C-301

BUN/UREA DT
Urea Nitrogen

INSTRUCTIONS FOR USE


Reagents

Reagents
Slide Diagram

Slide Ingredients
Reactive ingredients per cm
Urease flack beans, E.C.3.5.1.5) 1.2 U and N-propyl-4-(2,6-dinitro-4-chlorobenzyl)quinolonium ethane sulfonate (ammonia indicator) 0.26 mg.
; yA

Other ingredients
Pigment, binders, buffer, surfactants, stabilizers, chelator and cross-linking agent

1. Upper slide mount 2. Spreading layer (TIO2) 3. Reagent layer urease buffer.pH 7.8 4. Semipermeable membrane 5. Indicator layer ammonia indicator 6. Support layer 7. Lower slide mount

Slide Handling
CAUTION: Do not use slides with damaged or incompletely sealed packaging.

Slide Preparation
IMPORTANT: The slide must reach room temperature, 18 -28 C (64 -82 F), before the wrapper is opened. Do not use unopened slides that have been at room temperature, 18 "-28 C (64 -82 T) for >48 hours. 1. 2. Remove the unopened slide from the box. Warm the unopened slide for 15 minutes. Unopened slides that remain in the sealed wrapper may be returned to refrigerator or freezer storage. 3. Remove the packaging and place the slide into the loading station within 15 minutes after opening.

Slide Storage and Stability


VITROS BUN/UREA DT Slides are stable until the expiration date on the carton when they are stored and handled as specified.

Slide Storage and Stability for BUN/UREA DT


Slides Unopened Storage Condition Room temperature 18-28 'C (64-82 F) Refrigerated 2-8C (36-46F) Frozen <-18C (<0F) Room temperature 18-28 'C (64-82 F) Stability <48 hours Until expiration date Until expiration date <15 minutes

Opened

Specimen Requirements
WAR Mi HQ; ' Handle specimens as biohazardous material.

Specimens Recommended
| Serum Plasma:3 EDTA Meparin Certain collection devices have been reported to affect other analytes and tests.4 Confirm that your collection devices are compatible with this test.

IMPORTANT:

Specimens Not Recommended


Plasma:3 Sodium fluoride (fluoride inhibits the enzyme urease)

Serum and Plasma


Specimen Collection and Preparation
Collect specimens using standard laboratory procedures.5 6 Patient Preparation No special patient preparation is necessary.

Pub. No. C-301

Version 1.0

INSTRUCTIONS FOR USE


Testing Procedure

RUN/UREA OT
Urea Nitrogen

Special Precautions For the effect of sample hemolysis on test results, refer to "Limitations of the Procedure." Centrifuge serum and plasma specimens and remove the serum from the cellular material within 4 hours of collection.3 Specimen Handling and Storage

Handle specimens as biohazardous material. Handle and store specimens in stoppered containers to avoid contamination and evaporation. Mix samples by gentle inversion and bring to room temperature, 18-28C (64-82F), prior to analysis.

Specimen Storage and Stability for BUN/UREA DT: Serum and Plasma3 Storage Temperature Stability
Room temperature Refrigerated Frozen 18-28OC(64O-82F) 2-8C (36-46F) <-18C(<0F) <1 day <5 days <6 months

Testing Procedure
Materials Provided
. VITROS Chemistry Products BUN/UREA DT Slides

Materials Required But Not Provided


VITROS Chemistry Products DT Calibrator Kit Quality control materials, such as VITROS Chemistry Products DT Control I & II Isotonic saline or reagent-grade water VITROS DT Pipette

Operating Instructions
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System. Bring all fluids and samples to room temperature, 18-28C (64-82F), priorto analysis.

IMPORTANT;

Sample Dilution
If urea nitrogen concentrations exceed the system's reportable (dynamic) range: 1. Dilute the sample with isotonic saline or reagent-grade water. 2. Reanalyze. 3. Multiply the results by the dilution factor to obtain an estimate of the original sample's urea nitrogen concentration.

Calibration
Required Calibrators
VITROS Chemistry Products DT Calibrator Kit, bottles 1, 2, and 3

Calibrator Preparation, Handling, and Storage


Refer to the instructions for use for VITROS DT Calibrator Kit.

Calibration Procedure
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

When to Calibrate
Calibrate: When the slide lot number changes. When critical system parts are replaced due to service or maintenance. When government regulations require. - For example, in the USA, CLIA regulations require calibration or calibration verification at least once every six months.

Version 1.0

Pub. No. C-301

BUN/UREA DT
Urea Nitrogen

INSTRUCTIONS FOR USE


Quality Control

The VITROS BUN/UREA DT test may also need to be calibrated: If quality control results are consistently outside acceptable range. After certain service procedures have been performed. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Calculations
Reflectance from the slide is measured at 660 nm after the fixed incubation time. Once a calibration has been performed for each slide lot, urea nitrogen concentration in unknown samples can be determined using the software-resident endpoint colorimetric math model and the response obtained from each unknown test slide.

Validity of a Calibration
Calibration parameters are automatically assessed by the VITROS DT60/DT60 II Chemistry System against a set of quality parameters resident within the analyzer software. Failure to meet any of the pre-defined quality parameters results in a failed calibration. The calibration report should be used in conjunction with quality control results to determine the validity of a calibration.

Reportable (Dynamic) Range Reportable (Dynamic) Range for BUN/UREA DT


Conventional (mg/dL) 1-100 SI Units (mmol/L) 0.4-35.7

For out-of-range samples, refer to "Sample Dilution."

Traceability of the Calibration


Values assigned to the VITROS Chemistry Products DT Calibrator Kit for BUN/UREA are traceable to the Certified NIST (National Institute of Standards and Technology) Reference Material, SRM (Standard Reference Material) 912a. The Ortho-Clinical Diagnostics calibration laboratory uses SRM* 912a to calibrate the CDC Urease/GLDH method7 to support BUN/UREA value assignment for the VITROS DT Calibrator Kit.

Quality Control
Procedure Recommendations
| ' Handle quality control materials as biohazardous material. Choose control levels that check the clinically relevant range. Analyze quality control materials in the same manner as patient samples, before or during patient sample processing. To verify system performance, analyze control materials: - After calibration. - According to local regulations or at least once each day that the test is being performed. - After specified service procedures are performed. Refer to the operator's manual for your VITROS DT60/DT60II System. If control results fall outside your acceptable range, investigate the cause before deciding whether to report patient results. For general quality control recommendations, refer to Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition5 or other published guidelines. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Quality Control Material Selection


rftftiT: VITROS DT Control I & II are recommended for use with the VITROS DT60/DT60II Chemistry System. Evaluate the performance of other commercial control fluids for compatibility with this test before using for quality control.

Control materials other than VITROS DT Controls I & II may show a difference when compared with other urea nitrogen methods if they: - Depart from a true human matrix. - Contain high concentrations of preservatives, stabilizers, or other nonphysiological additives. Do not use control materials stabilized with ethylene glycol. Some controls that are low in carbon dioxide concentration may show a negative bias (>10% at CO2 <8 mmol/L) that may be avoided by reconstituting lyophilates with a bicarbonate diluent instead of with water. Ammonium bicarbonate diluent should not be used as it will cause a positive bias in test results.

Pub. No. C-301

Version 1.0

[S| VITR!

INSTRUCTIONS FOR USE


Expected Values and Reporting Units

BUN/UREA OT
Urea Nitrogen

Proficiency survey samples may show a negative bias similar to controls low in CO2. Contact the testing agency for instructions because reconstituting with special diluents may affect other analyte values (e.g., reconstituting with sodium bicarbonate will affect sodium proficiency scores).

Quality Control Material Preparation and Storage


Refer to the instructions for use for VITROS Chemistry Products DT Control I & II or to other manufacturer's product literature.

Expected Values and Reporting Units


Reference Interval
The serum reference interval is the central 95% of results from an internal study of 3160 apparently healthy adults from a working population (612 females and 2548 males).

Reference Interval for BUN/UREA DT


Conventional Units (mg/dL) Male Female

9-20 7-17

SI Units (mmol/L) 3.2-7.1 2.5-6.1

Each laboratory should confirm the validity of these intervals for the population it serves.

Reporting Units and Unit Conversion


The VITROS DT60/DT60 II Chemistry System may be programmed to report urea nitrogen results in conventional and SI units.

Reporting Units and Unit Conversion for BUN/UREA DT Conventional Units SI Units
mg/dL urea N mmol/L urea (mg/dL urea N x 0.3569)

Limitations of the Procedure


Known Interferences
Ammonium ions may cause an increase in measured BUN/UREA value equivalent to the specimen's nitrogen content.9 The VITROS BUN/UREA DT Slide method was screened for interfering substances following NCCLS Protocol EP7.10 The substances listed in the table, when tested at the concentrations indicated, caused the bias shown.

Known Interfering Substances for BUN/UREA DT


Interferent* Hemoglobin
*

Interferent Concentration 50 mg/dL (0.5 g/L)

Blood Urea Nitrogen Concentration Conv. (mg/dL) SI (mmol/L) 28 10

Bias Conv. (mg/dL) SI (mmol/L)


1.1 0.4

It is possible that other interfering substances may be encountered. These results are representative; however, your results may differ somewhat due to test-to-test variation. The degree of interference at concentrations other than those listed might not be predictable.

Other Limitations
Certain drugs and clinical conditions are known to alter blood urea nitrogen concentration in vivo. For additional information, refer to one of the published summaries.11 12

Version 1.0

Pub. No. C-301

BUN/UREA DT
Urea Nitrogen

INSTRUCTIONS FOR USE


Performance Characteristics

Performance Characteristics
Method Comparison

The plots and table show the results of a comparison of samples analyzed on the VITROS DT60 II System with those analyzed using the VITROS 950 System. Testing followed NCCLS Protocol EP9. Method Comparison for BUN/UREA DT: Serum Conventional Units 120 3
Ol

SI Units
40

y =x
o E

100
80 60

3(1

20

40 10 20 0

20

40

60

100

120

10

20

30

40

Comparative Method: VITROS 950 System (mg/dL)

Comparative Method. VITROS 950 System (mmol/L)

Method Comparison for BUN/UREA DT: Serum


Conventional Units (mg/dL) n DT60 II System vs. 950 System 62 Correlation Slope Coefficient 0.98 0.998 Range of Sample Cone. 5-89 Intercept -0.10 Sy.x 1.61 SI Units (mmol/L) Range of Sample Cone. 1.8-31.7 Intercept -0.03 Sy.x 0.58

Precision
Precision was evaluated with quality control materials on the VITROS DT60 II System following NCCLS Protocol EP5.14 The data presented are a representation of test performance and are provided as a guideline. Variables such as sample handling and storage, reagent handling and storage, laboratory environment, and system maintenance can affect reproducibility of test results.

Precision for BUN/UREA DT: Serum


Conventional Units (mg/dL) System VITROS DT60 II Mean Cone. 18 51 Within Day SD* 0.5 1.4 Within Lab SD** 0.7 1.9 SI Units (mmol/L) Mean Cone. 6.3 18.4 Within Day SD* 0.18 0.49 Within Lab SD** 0.26 0.69 Within Lab CV%** 4.1 3.7 No. Observ. 88 88 No. Days 22 22

Within Day precision was determined using two runs/day with two replications. Within Lab precision was determined using a single lot of slides and calibrating weekly.

Pub. No. C-301

Version 1.0

INSTRUCTIONS FOR USE


References

BUN/UREA DT
Urea Nitrogen

References
1. 2. 3. 4. 5. 6. 7. Tietz NW (ed). Fundamentals of Clinical Chemistry, ed. 3. Philadelphia: WB Saunders; 967; 1987. NCCLS. Protection of Laboratory Workers from Instrument Biohazards and Infectious Diseases Transmitted by Blood, Body Fluids and Tissue; Approved Guideline. NCCLS Document M29 (OSBN 1-56238). NCCLS, Wayne, PA 19087; 1997. Clinical Laboratory Handbook for Patient Preparation and Specimen Handling. Fascicle VI: Chemistry/Clinical Microscopy. Northfield, IL: College of American Pathologists; 1992. Calam RR. Specimen Processing Separator Gels: An Update. J Clin Immunoassay. 11:86-90; 1988. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Venipuncture. NCCLS Document H3. Wayne, PA: NCCLS;

1991.
NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Skin Puncture. NCCLS Document H4. Wayne, PA: NCCLS;

1991.
Sampson Rl, et al. A coupled-enzyme equilibrium method for measuring urea in serum: optimization and evaluation of the AACC study group on urea candidate reference method. Clin. Chem. 26:816-26; 1980. 8. NCCLS. Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition. NCCLS Document C24. Wayne, PA: NCCLS; 1999. 9. Tietz NW(ed). Fundamentals of Clinical Chemistry, ed. 3. Philadelphia: WB Saunders; 676-679; 1987. 10. NCCLS. Interference Testing in Clinical Chemistry. NCCLS Document EP7. Wayne, PA: NCCLS; 1986. 11. Young DS. Effects of Drugs on Clinical Laboratory Tests, ed. 4. Washington D C : AACC Press; 1995. 12. Friedman RB, Young DS. Effects of Disease on Clinical Laboratory Tests. Washington, D.C.: AACC Press; 1990. 13. NCCLS. Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline. NCCLS Document EP9. Wayne, PA: NCCLS; 1995. 14. NCCLS. User Evaluation of Precision Performance with Clinical Chemistry Devices. NCCLS Document EP5. Wayne, PA: NCCLS;

1992.

Glossary of Symbols
The Glossary of Symbols table defines the symbols used on the VITROS Chemistry Products packaging and on the VITROS Chemistry Products Assay Sheets.

Glossary of Symbols
Do Not Reuse Use by or Expiration Date (YYYY-MM-DD) Lot Number Manufacturer's Serial Number Catalog Number or Product Code Attention: See Instructions for Use. Manufacturer Authorized Representative Contains Sufficient for "n" Tests For In Vitro Diagnostic Use Store Between Consult Instructions for Use Fragile, Handle with Care. Keep Dry This end up

Store At or Below Store At or Above

Version 1.0

Pub. No. C-301

VITFJCpS H

BUN/UREA DI
Urea Nitrogen

INSTRUCTIONS FOR USE


Revision History

Revision History
Date of Revision 2003-10-01 Version 1.0 Description of Technical Changes* New format > New organization and sections consistent with IVD Directive Specimens Recommended - plasma: added EDTA Specimen Storage and Stability - updated stability values > Quality Control Material Selection - added data > Method Comparison - updated all comparisons and the plot > Precision - updated all data References - added all

The change bars indicate the position of a technical amendment to the text with respect to the previous version of the document.

When this Instructions For Use is replaced, sign and date below and retain as specified by local regulations or laboratory policies, as appropriate.

Signature

Obsolete Date

C
I EC I REP I
Ortho-Clinical Diagnostics Mandeville House 62 The Broadway Amersham Buckinghamshire HP7 OHJ England Ortho-Clinical Diagnostics, Inc. 100 Indigo Creek Drive Rochester, NY 14626-5101

' Ortho-Clinical Diagnostics


company

VITROS is a trademark of Ortho-Clinical Diagnostics, Inc. Ortho-Clinical Diagnostics, Inc., 2003. All rights reserved. Printed in USA.

Pub.

No. C-301

Version 1.0

Products

Chemistry

INSTRUCTIONS FOR USE


VITROS Chemistry Products Ca DT Slides Intended Use
For in vitro diagnostic use only. VITROS Ca DT Slides qualitatively measure calcium (Ca) concentration in serum and plasma.

CaOT
Calcium

Summary and Explanation of the Test


Calcium is the major mineral component of bone; 99% of the body's calcium is in bone. Calcium ions play an important role in the transmission of nerve impulses and in maintaining normal muscle contraction. Abnormal concentrations of serum calcium may indicate malfunction of the parathyroid glands, bone diseases, carcinoma, malnutrition and malabsorption syndrome, vitamin D deficiency, overdose with calcium-containing antacids, and renal diseases.1

Principles of the Procedure


The VITROS Ca DT Slide method is performed using the VITROS Ca DT Slide and the VITROS Chemistry Products DT Calibrator Kit on VITROS DT60/DT60 II Chemistry Systems. The VITROS Ca DT Slide is a multilayered, analytical element coated on a polyester support. A drop of patient sample is deposited on the slide and is evenly distributed by the spreading layer to the underlying layers. The bound calcium is dissociated from binding proteins, allowing the calcium to penetrate through the spreading layer into the underlying reagent layer. There, the calcium forms a complex with Arsenazo III dye, causing a shift in the absorption maximum. After incubation, the reflection density of the colored complex is measured spectrophotometrically. The amount of colored complex formed is proportional to the calcium concentration in the sample.

Reaction Sequence
Ca+2 + Arsenazo I
pH5.6

colored complex

Test Type and Conditions


Test Type and Conditions for Ca DT
Test Type Colorimetric VITROS DT60/DT60 II Module DTSC/DTSC II Approximate Incubation Time 5 minutes Temperature 37C (98.6F) Wavelength 680 nm Sample Drop Volume 10 pL

Warnings and Precautions


For in vitro diagnostic use only. Take care when handling materials and samples of human origin. Since no test method can offer complete assurance that infectious agents are absent, consider all clinical specimens, controls, and calibrators potentially infectious. Handle specimens, solid and liquid waste, and test components in accordance with local regulations and NCCLS Guideline M292 or other published biohazard safety guidelines. For specific warnings and precautions for calibrators, quality control materials, and other components, refer to the instructions for use for the appropriate VITROS product, or to other manufacturer's product literature.

Version 1.0

Pub. No. C-348

VITRJJD5 Q

CaDT
Calcium

INSTRUCTIONS FOR USE


Reagents

Reagents
Slide Diagram

Slide Ingredients |
|

1 . 2 .3

Reactive ingredients per cm2 Arsenazo III dye 60 |xg.


Other ingredients Pigment, binders, surfactants, buffer, cross-linking agent and mordant.
~~ "
. . ' -

1. Upper slide mount 2. Spreading layer (TIO2)


3. Reagent layer Arsenazo III dye buffer, pH 5.6 4. Support layer S. Lower slide mount

~4
'

Slide Handling
CAUTION: Do not use slides with damaged or incompletely sealed packaging.

Slide Preparation
The slide must reach room temperature, 18-28C (64-82F), before the wrapper is opened. Do not use unopened slides that have been at room temperature, 18-28C (64-82F) for >48 hours. 1. 2. Remove the unopened slide from the box. Warm the unopened slide for 15 minutes. Unopened slides that remain in the sealed wrapper may be returned to refrigerator or freezer storage. 3. Remove the packaging and place the slide into the loading station within 15 minutes after opening.

Slide Storage and Stability


VITROS Ca DT Slides are stable until the expiration date on the carton when they are stored and handled as specified.

Slide Storage and Stability for Ca DT


Slides Unopened Opened

Storage Condition Room temperature 18-28C(64-82 F) Refrigerated 2-8C (36-46F) Frozen <-18C (<0F) Room temperature 18-28C(64-82 F)

Stability <48 hours Until expiration date Until expiration date <15 minutes

Specimen Requirements
Handle specimens as biohazardous material.
CAUTION: Protective gloves manufactured with calcium carbonate powders may cause elevated test results because of the contamination of sample handling supplies (for example, pipette tips, transfer pipettes, sample cups and caps). Supplies that have come in contact with powdered gloves may subsequently contaminate the test specimen during sample metering. Gloves labeled as "powder-free" may contain some contaminating powder agents on the inside of the glove.

NOTE:

Pub. No. C-348

Version 1.0

INSTRUCTIONS FOR USE


Testing Procedure Specimens Recommended
Serum Plasma:3 Heparin Certain collection devices have been reported to affect other analytes and tests4 Confirm that your collection devices are compatible with this test.

Ca DT
Calcium

IMPORTANT:

Specimens Not Recommended


Plasma:5 EDTA* Fluoride oxalate* Citrate* "These substances chelate calcium, causing negative bias.5 Do not use blood from patients on EDTA therapy.

Specimen Collection and Preparation


Collect specimens using standard laboratory procedures.6 7 Patient Preparation No special patient preparation is necessary. Special Precautions Results from recumbent patients may be 3% lower.8 Blood collected with stasis may have calcium concentrations 15% higher.8 Centrifuge specimens and remove the serum and plasma from the cellular material within 2 days of collection.9

Specimen Handling and Storage


Handle and store specimens in stoppered containers to avoid contamination and evaporation. Mix samples by gentle inversion and bring to room temperature, 18-28C (64-82F), prior to analysis.

Specimen Storage and Stability for Ca DT: Serum and Plasma


Storage Room temperature Refrigerated9 Frozen9 Temperature 18-28OC(64O-82F) 2-8C (36-46F) <-18C(<0F) Stability <4 hours <22 days <1 year

Testing Procedure
Materials Provided
. VITROS Chemistry Products Ca DT Slides

Materials Required But Not Provided


. VITROS Chemistry Products DT Calibrator Kit Quality control materials, such as VITROS Chemistry Products DT Control I & II Isotonic saline Reagent-grade water VITROS DT Pipette

Operating Instructions
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System. Bring all fluids and samples to room temperature, 18-28C (64-82F), prior to analysis.

Version 1.0

Pub. No. C-348

VITFJCpIS 0

CaDT
Calcium

INSTRUCTIONS FOR USE


Calibration
If calcium concentrations exceed the system's reportable (dynamic) range: 1. Dilute with isotonic saline or reagent-grade water. 2. Reanalyze. 3. Multiply the results by the dilution factor to obtain an estimate of the original sample's calcium concentration.

Sample Dilution

Calibration
Required Calibrators
VITROS Chemistry Products DT Calibrator Kit, bottles 1, 2, and 3

Calibrator Preparation, Handling, and Storage


Refer to the instructions for use for VITROS DT Calibrator Kit.

Special Precautions
If the laboratory's ambient temperature has changed 5F (3C), or more, from the temperature at the time that the calcium test was calibrated, then the quality-control materials should be checked. If the quality-control materials are out of control, recalibrate the analyzer for calcium and record the temperature at the time of calibration for future reference.

Calibration Procedure
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

When to Calibrate
Calibrate: When the slide lot number changes. When critical system parts are replaced due to service or maintenance. When government regulations require. - For example, in the USA, CLIA regulations require calibration or calibration verification at least once every six months. The VITROS Ca DT test may also need to be calibrated: If quality control results are consistently outside acceptable range. After certain service procedures have been performed. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Calculations

I
I

Reflectance from the slide is measured at 680 nm after the fixed incubation time. Once a calibration has been performed for each slide lot, calcium concentration in unknown samples can be determined using the software-resident endpoint colorimetric math model and the response obtained from each unknown test slide.

Validity of a Calibration
Calibration parameters are automatically assessed by the VITROS DT60/DT60 II Chemistry System against a set of quality parameters resident within the analyzer software. The quality control results should be used to determine the validity of a calibration.

Reportable (Dynamic) Range Reportable (Dynamic) Range for Ca DT


Serum Conventional (mg/dL) 3.00-14.00 SI Units (mmol/L) 0.75-3.49

For out-of-range samples, refer to "Sample Dilution."

Traceability of the Calibration


Values assigned to the VITROS Chemistry Products DT Calibrator Kit for calcium are traceable to the Certified NIST (National Institute of Standards and Technology) Reference Material, SRM (Standard Reference Material) 915a. The Ortho-Clinical Diagnostics calibration laboratory uses SRM 915a to calibrate the Flame Atomic Absorption Spectroscopy method10 to support calcium value assignment for the VITROS Chemistry Products DT Calibrator Kit.

Pub. No. C-348

Version 1.0

INSTRUCTIONS FOR USE


Quality Control

Ca DT
Calcium

Quality Control
Procedure Recommendations
I Handle quality control materials as biohazardous material. Choose control levels that check the clinically relevant range. Analyze quality control materials in the same manner as patient samples, before or during patient sample processing. To verify system performance, analyze control materials: - After calibration. - According to local regulations or at least once each day that the test is being performed. - After specified service procedures are performed. Refer to the operator's manual for your VITROS DT System. If control results fall outside your acceptable range, investigate the cause before deciding whether to report patient results. For general quality control recommendations, refer to Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition" or other published guidelines. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Quality Control Material Selection

IMPORTANT:

VITROS DT Control I & II are recommended for use with the VITROS DT60/DT60 II

Chemistry System. Evaluate the performance of other commercial control fluids for compatibility with this test before using for quality control. Control materials other than VITROS DT Controls I & II may show a difference when compared with other calcium methods if they: - Depart from a true human matrix. - Contain high concentrations of preservatives, stabilizers, or other nonphysiological additives. Do not use control materials stabilized with ethylene glycol.

Quality Control Material Preparation and Storage


Refer to the instructions for use for VITROS Chemistry Products DT Control I & II or to other manufacturer's product literature.

Expected Values and Reporting Units


Reference Interval
These reference intervals are based on an external study.12

Reference Interval for Ca DT


Conventional Units (mg/dL) 8.4-10.2

SI Units (mmol/L) 2.10-2.55

Each laboratory should confirm the validity of these intervals for the population it serves.

Reporting Units and Unit Conversion


The VITROS DT60/DT60 II Chemistry System may be programmed to report calcium results in conventional and SI units.

Reporting Units and Unit Conversion for Ca DT


Conventional Units mg/dL

SI Units mmol/L (mg/dL x 0.2495)

Version 1.0

Pub. No. C-348

CaDT
Calcium

INSTRUCTIONS FOR USE


Limitations of the Procedure

Limitations of the Procedure


Known Interferences
* Blood from patients receiving Hypaque radiographic contrast agent cannot be used. Suramin, an antiparasitic drug, has been reported to cause a bias of -10% in calcium results at a suramin concentration of 300 ug/mL.13

Other Limitations
Keeping the sample in an open container at room temperature may increase the reported calcium concentration by up to 0.4 mg/dL (0.1 mmol/L). Changes are due to the loss of carbon dioxide, which results in an increase in pH of the specimen. The increase is minimized by anaerobic handling procedures and prompt analysis. Adherence to these procedures is especially important for pediatric samples where the sample volume is small. Certain drugs and clinical conditions are known to alter calcium concentration in vivo. For additional information, refer to one of the published summaries.1'1'15

Performance Characteristics
Method Comparison
The plots and tables show the results of a comparison of samples analyzed on the VITROS DT60 II System with those analyzed using the Atomic Absorption comparative method.10 Testing followed NCCLS Protocol EP9.16

Method Comparison for Ca DT: Serum


Conventional Units
15

SI Units
y =x

y =x

12

I
9 6

CO

3 0

8 o
t
15
Comparative Method: Atomic Absorption (mg/dL)

Comparative Method: Atomic Absorption (mmol/L)

Method Comparison for Ca DT: Serum


Conventional Units or (mg/dL) n DT60 II System vs. comparative method 77 Correlation Slope Coefficient 0.98 0.997 Range of Sample Cone. Intercept Sy.x 0.21 0.20 SI Units (mmol/L) Range of Sample Cone. Intercept 0.05 Sy.x 0.05

3.0-12.9

0.76-3.21

Pub. No. C-348

Version 1.0

{I VITRJ

INSTRUCTIONS FOR USE


References Precision

CaDT
Calcium

Precision was evaluated with quality control materials on the VITROS DT60 II System following NCCLS Protocol EP5.17 The data presented are a representation of test performance and are provided as a guideline. Variables such as sample handling and storage, reagent handling and storage, laboratory environment, and system maintenance can affect reproducibility of test results. Precision for Ca DT: Serum Conventional Units (mg/dL) System DT60 II System 11.4 0.09 0.21 2.85 0.02 0.05 1.8 87 22 Within Day precision was determined using two runs/day with two replications. Within Lab precision was determined using a single lot of slides and calibrating weekly. Mean Cone. 9.1 Within Day SD* 0.10 Within Lab SD** 0.18 SI Units (mmol/L) Mean Cone. 2.26 Within Day SD* 0.03 Within Lab SD** 0.04 Within Lab CV%** 2.0 No. Observ. 88 No. Days 22

References
1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16. 17. Tietz NW(ed). Fundamentals of Clinical Chemistry, ed. 3. Philadelphia: WB Saunders; 705-713; 1987. NCCLS. Protection of Laboratory Workers from Instrument Biohazards and Infectious Diseases Transmitted by Blood, Body Fluids and Tissue; Approved Guideline. NCCLS Document M29 (OSBN 1-56238). NCCLS, Wayne, PA 19087; 1997. Doumas BT, et al. Differences Between Values for Plasma and Serum in Tests Performed in the Ektachem 700 XR Analyzer, and Evaluation of "Plasma Separator Tubes (PST)." Clin. Chem. 35:151-153; 1989. Calam RR. Specimen Processing Separator Gels: An Update. J Clin Immunoassay. 11:86-90; 1988. Tietz NW. Textbook of Clinical Chemistry, ed. 2. Philadelphia: WB Saunders; 66-67,1900; 1994. NCCLS. Procedures forthe Collection of Diagnostic Blood Specimens by Venipuncture. NCCLS Document H3. Wayne, PA: NCCLS; 1991. NCCLS. Procedures forthe Collection of Diagnostic Blood Specimens by Skin Puncture. NCCLS Document H4. Wayne, PA: NCCLS; 1991. Tietz NW. Textbook of Clinical Chemistry, ed. 2. Philadelphia: WB Saunders; 60, 80; 1994. Clinical Laboratory Handbook for Patient Preparation and Specimen Handling. Fascicle VI: Chemistry/Clinical Microscopy. Northfield, IL: College of American Pathologists; 1992. Cali JP, et al. Atomic Absorption. NBS Reference Method (modified). Clin. Chem. 19:1208; 1987. NCCLS. Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition. NCCLS Document C24. Wayne, PA: NCCLS; 1999. Tietz NW(ed). Fundamentals of Clinical Chemistry, ed. 3. Philadelphia: WB Saunders; 947; 1987. Gregory, et al. Suramin Interferes with Measurements of Total Calcium and Serum Amylase by the Kodak Ektachem 700 Analyzer and May Inhibit Liver Enzyme Activity. Clin. Chem. 38:2552-2553; 1992. Young DS. Effects of Drugs on Clinical Laboratory Tests, ed. 4. Washington D C : AACC Press; 1995. Friedman RB, Young DS. Effects of Disease on Clinical Laboratory Tests. Washington, D.C.: AACC Press; 1990. NCCLS. Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline. NCCLS Document EP9. Wayne, PA: NCCLS; 1995. NCCLS. User Evaluation of Precision Performance with Clinical Chemistry Devices. NCCLS Document EP5. Wayne, PA: NCCLS; 1992.

Version 1.0

Pub. No. C-348

VITRCpS 0

CaDT
Calcium

INSTRUCTIONS FOR USE


Glossary of Symbols

Glossary of Symbols
The Glossary of Symbols table defines the symbols used on the VITROS Chemistry Products packaging and on the VITROS Chemistry Products Assay Sheets.

Glossary of Symbols
Q<J Do Not Reuse
Use by or Expiration Date (YYYY-MM-DD) Lot Number Manufacturer's Serial Number Catalog Number or Product Code Attention: See Instructions for Use. EC I REP I Manufacturer Auth'orized Representative Contains Sufficient for "n" Tests For In Vitro Diagnostic Use Store At or Below Store At or Above

2
| LOT | QKI OlN

m !

Store Between Consult Instructions for Use Fragile, Handle with Care. Keep Dry This end up

REF

Revision History
Date of Revision 2003-03-28 Version 1.0 Description of Technical Changes* New organization and sections consistent with IVD Directive Specimen Handling and Storage - updated stabilities; removed "Serum should not be frozen." Specimen Collection and Preparation - updated all statements under Special Precautions Limitations of the Procedure - added suramin Method Comparison - updated comparison data and plots Precision - updated all data References - added 2, 3, 4, 5, 7, 8, 9, 10, 11, 12, 13, 16, 17

The change bars indicate the position of a technical amendment to the text with respect to the previous version of the document.

When this Instructions For Use is replaced, sign and date below and retain as specified by local regulations or laboratory policies, as appropriate.

Signature

Obsolete Date

C
Ortho-Clinical Diagnostics Mandeville House 62 The Broadway Amersham Buckinghamshire HP7 OHJ England Ortho-Clinical Diagnostics, Inc. 100 Indigo Creek Drive Rochester, NY 14626-5101

' Ortho-Clinical Diagnostics


8H company

VITROS is a trademark of Ortho-Clinical Diagnostics, Inc. Ortho-Clinical Diagnostics, Inc., 2003. All rights reserved. Printed in USA.

Pub. No. C-348

Version 1.0

viTRras
Chemistry!

INSTRUCTIONS FOR USE


VITROS Chemistry Products CHE DT Slides Intended Use
For in vitro diagnostic use only. VITROS CHE DT Slides quantitatively measure cholinesterase (CHE) activity in serum and plasma.

CHEOT
Cholinesterase

Summary and Explanation of the Test


There are two types of cholinesterase: Acetylcholinesterase (EC.3.1.1.7), which is found in red blood cells and nerve tissues. Cholinesterase (E.C.3.1.1.8), which is found in plasma, liver, heart, and other tissues. These measurements are useful in the diagnosis of pesticide poisoning, liver diseases and sensitivity to succinylcholine administration.12 Pesticide poisoning. Organophosphate and carbamate pesticides are inhibitors of both cholinesterase and acetylcholinesterase. Although the toxic effect is caused by inhibition of acetylcholinesterase in nerve endings, cholinesterase is often used clinically because it is present in serum in high activities and is easy to measure. Liver diseases. Cirrhosis, hepatitis, and carcinoma with metastasis to the liver are known to lower cholinesterase activity. A decrease in CHE activity is considered a sensitive measure of a drop in liver synthetic capacity, because high activities of cholinesterase are normally present in serum. Sensitivity to succinylcholine administration. Succinylcholine is a short-acting muscle relaxant administered during surgery. It is a reversible inhibitor of acetylcholinesterase and is hydrolyzed by serum cholinesterase. Individuals without sufficient serum cholinesterase activity or with certain genetic variants may be unable to metabolize the drug quickly, resulting in prolonged apnea. Low CHE activities may be chronic for the individual or transient due to pesticide exposure, liver disorder, pregnancy, or the use of oral contraceptives.

Principles of the Procedure


The VITROS CHE DT Slide method is performed using the VITROS CHE DT Slide and the VITROS Chemistry Products DT Specialty Calibrator Kit on VITROS DT60/DT60 II Chemistry Systems. The VITROS CHE DT Slide is a multilayered, analytical element coated on a polyester support. A drop of patient sample is deposited on the slide and is evenly distributed by the spreading layer to the underlying layers. Cholinesterase hydrolyzes butyrylthiocholine to thiocholine. The liberated thiocholine reduces potassium hexacyanoferrate III (potassium ferricyanide) to potassium hexacyanoferrate II (potassium ferrocyanide). The rate of color loss is monitored by reflectance spectrophotometry. The rate of change in reflection density is proportional to the cholinesterase activity in the sample.

Reaction Sequence
butyrylthiocholine + H2O
CHE

thiocholine + butyrate dithiobis(choline) + 2 potassium ferrocyanide

2 thiocholine + 2 potassium ferricyanide

Test Type and Conditions


Test Type and Conditions for CHE DT
Test Type Rate VITROS DT60/DT60 II Module DTSC/DTSC II Approximate Incubation Time 5 minutes Temperature 37C (98.6F) Wavelength 400 nm Sample Drop Volume 10 ML

Version 1.0

Pub. No. C-358

CHEDT
Cholinesterase

INSTRUCTIONS FOR USE


Warnings and Precautions

Warnings and Precautions


For in vitro diagnostic use only. Take care when handling materials and samples of human origin. Since no test method can offer complete assurance that infectious agents are absent, consider all clinical specimens, controls, and calibrators potentially infectious. Handle specimens, solid and liquid waste, and test components in accordance with local regulations and NCCLS Guideline M293 or other published biohazard safety guidelines. For specific warnings and precautions for calibrators, quality control materials, and other components, refer to the instructions for use for the appropriate VITROS product, or to other manufacturer's product literature.

Reagents
Slide Diagram

Slide Ingredients
I Reactive ingredients per cm Potassium ferricyanide 180 ug and butyrylthiocholine iodide 290 ug.
2

y'

..-

Other ingredients
Pigment, binders, buffer, surfactants and cross-linking agent.

1. Upper slide mount 2. Spreading layer (TIO2) butyrylthiocholine iodide 3. Reagent layer potassium ferricyanide buffer, pH 7.6 4. Support layer 5. Lower slide mount

Slide Handling
CAUTSQN:

Do not use slides with damaged or incompletely sealed packaging.

Slide Preparation
IMPORTANT: The slide must reach room temperature, 18"-28 C (64 -82 F), before the wrapper is opened. Do not use unopened slides that have been at room temperature, 18 -28 C (64 -82 F) for >48 hours. 1. 2. Remove the unopened slide from the box. Warm the unopened slide for 15 minutes. Unopened slides that remain in the sealed wrapper may be returned to refrigerator or freezer storage. 3. Remove the packaging and place the slide into the loading station within 15 minutes after opening.

Slide Storage and Stability


VITROS CHE DT Slides are stable until the expiration date on the carton when they are stored and handled as specified.

Slide Storage and Stability for CHE DT


Slides Unopened Opened

Storage Condition Room temperature 18-28C (64-82F) Frozen <-18C(<0F) Room temperature 18-28C (64-82F)

Stability <48 hours Until expiration date <15 minutes

Specimen Requirements
WA R H! N S: Handle specimens as biohazardous material.

Specimens Recommended
I I Serum Plasma: Heparin Certain collection devices have been reported to affect other anaiytes and tests.4 Confirm that your collection devices are compatible with this test.

IMPORTANT:

Specimens Not Recommended


Do not use visibly hemolyzed specimens.5

Pub. No. C-358

Version 1.0

[J V I T R I

INSTRUCTIONS FOR USE


Testing Procedure Serum and Plasma Specimen Collection and Preparation
Collect specimens using standard laboratory procedures. 6 ' Patient Preparation No special patient preparation is necessary. Special Precautions Centrifuge specimens and remove the serum from the cellular material within 4 hours of collection.8

CHEDT
Cholinesterase

Specimen Handling and Storage 41N6: Handle specimens as biohazardous material.

Handle and store specimens in stoppered containers to avoid contamination and evaporation. Mix samples by gentle inversion and bring to room temperature, 18-28C (64-82F), prior to analysis. Specimen Storage and Stability for CHE DT: Serum and Plasma8 Storage Room temperature Refrigerated Frozen Temperature 18-28C (64-82F) 2-8C (36-46F) <-18C(<0F) Stability <6 hours <7 days Not recommended

Testing Procedure
Materials Provided
VITROS Chemistry Products CHE DT Slides

Materials Required But Not Provided


I VITROS Chemistry Products DT Specialty Calibrator Kit Quality control materials, such as VITROS Chemistry Products DT Control I & II VITROS Chemistry Products 7% BSA or isotonic saline VITROS DT Pipette

Operating Instructions
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System. Bring all fluids and samples to room temperature, 18-28C (64-82F), priorto analysis. IMPORTANT:

Sample Dilution
I If cholinesterase activities exceed the system's reportable (dynamic) range: 1. Dilute the sample with VITROS 7% BSA or isotonic saline. 2. Reanalyze. 3. Multiply the results by the dilution factor to obtain an estimate of the original sample's cholinesterase activity.

Calibration
Required Calibrators
VITROS Chemistry Products DT Specialty Calibrator Kit, bottles 1, 2, and 4

Calibrator Preparation, Handling, and Storage


Refer to the instructions for use for VITROS DT Specialty Calibrator Kit.

Calibration Procedure
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Version 1.0

Pub. No. C-358

GHEDT
Cholinesterase

INSTRUCTIONS FOR USE


Quality Control

When to Calibrate
Calibrate: When the slide lot number changes. When critical system parts are replaced due to service or maintenance. When government regulations require. - For example, in the USA, CLIA regulations require calibration or calibration verification at least once every six months. The VITROS CHE DT test may also need to be calibrated: If quality control results are consistently outside acceptable range. After certain service procedures have been performed. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Calculations
Based on sequential readings of the slide's reflectance at 400 nm over the defined incubation period, a rate of change in reflectance is determined. This rate is used in the software-resident multi-point rate calibration model to compute enzyme activity. Once a calibration has been performed for each slide lot, cholinesterase activity in unknown samples can be determined from the rate of change in reflectance measured for each unknown test slide.

Validity of a Calibration
Calibration parameters are automatically assessed by the VITROS DT60/DT60 II Chemistry System against a set of quality parameters resident within the analyzer software. Failure to meet any of the pre-defined quality parameters results in a failed calibration. The calibration report should be used in conjunction with quality control results to determine the validity of a calibration.

Reportable (Dynamic) Range Reportable (Dynamic) Range for CHE DT


Conventional (U/mL) SI Units (U/L) 200-12500

0.20-12.50

For out-of-range samples, refer to "Sample Dilution."

Traceability of the Calibration

I
|

Values assigned to the VITROS Chemistry Products DT Specialty Calibrator Kit for cholinesterase are traceable to the butyrylthiocholine-based ferricyanide cholinesterase method recommended by the German Society for Clinical Chemistry,9 measured on a centrifugal analyzer at 37C.

Quality Control
Procedure Recommendations
WARMING: Handle quality control materials as biohazardous material. Choose control levels that check the clinically relevant range. Analyze quality control materials in the same manner as patient samples, before or during patient sample processing. To verify system performance, analyze control materials: - After calibration. - According to local regulations or at least once each day that the test is being performed. - After specified service procedures are performed. Refer to the operator's manual for your VITROS DT60/DT60 II System. If control results fall outside your acceptable range, investigate the cause before deciding whether to report patient results. For general quality control recommendations, refer to Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition10 or other published guidelines. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Pub.

No. C-358

Version 1.0

INSTRUCTIONS FOR USE


Expected Values and Reporting Units

CHEDT
Cholinesterase

Quality Control Material Selection


IMP
VITROS DT Control I & II are recommended for use with the VITROS DT60/DT60 II Chemistry System. Evaluate the performance of other commercial control fluids for compatibility with this test before using for quality control.

Control materials other than VITROS DT Controls I & II may show a difference when compared with other cholinesterase methods if they: - Depart from a true human matrix. - Contain high concentrations of preservatives, stabilizers, or other nonphysiological additives. Do not use control materials stabilized with ethylene glycol.

Quality Control Material Preparation and Storage


Refer to the instructions for use for VITROS Chemistry Products DT Control I & II or to other manufacturer's product literature.

Expected Values and Reporting Units


Reference Interval
These reference intervals are the central 95% of results from an internal study of 240 apparently healthy adults from a working population (101 females and 139 males).

Reference Interval for CHE DT


Conventional Units (U/mL) 5.90-12.22 4.65-10.44 SI Units (U/L) 5900-12220 4650-10440

Male Female

Each laboratory should confirm the validity of these intervals for the population it serves.

Reporting Units and Unit Conversion


The VITROS DT60/DT60 II Chemistry System may be programmed to report cholinesterase results in conventional and SI units.

Reporting Units and Unit Conversion for CHE DT Conventional Units SI Units
U/mL U/L (U/mL x 1000)

Limitations of the Procedure


Known Interferences
The VITROS CHE DT Slide method was screened for interfering substances following NCCLS Protocol EP7.11 The substances listed in the table, when tested at the concentrations indicated, caused the bias shown.

Known Interfering Substances for CHE DT


Interferent* Procainamide Phenazopyridine L-dopa
*

Interferent Concentration 4 mg/mL 10 mg/mL 80 ug/dL 80 ng/dL 300 (.ig/mL 17 mmol/L 42 mmol/L 3.2 (.imol/L 3.2 |.imol/L 1.5 mmol/L

Cholinesterase Activity Conv. (U/mL) SI (U/L) 6.5 6.5 4.5 6.5 6.0 6500 6500 4500 6500 6000

Bias Conv. (U/mL) SI (U/L) -0.48 -1.05 -0.74 -0.73 -1.11 -477 -1050 -740 -730 -1114

It is possible that other interfering substances may be encountered. These results are representative; however, your results may differ somewhat due to test-to-test variation. The degree of interference at concentrations other than those listed might not be predictable.

Other Limitations
Low pH (6.8) causes a 15% negative bias. Certain drugs and clinical conditions are known to alter cholinesterase activity in vivo. For additional information, refer to one of the published summaries.12 13

Version 1.0

Pub. No. C-358

GHEDI
Cholinesterase

INSTRUCTIONS FOR USE


Performance Characteristics

Performance Characteristics
Method Comparison

The plots and table show the results of a comparison of samples analyzed on the VITROS DT60 II System with those analyzed using the VITROS 950 System. Testing followed NCCLS Protocol EP9.14 Method Comparison for CHE DT: Serum Conventional Units 15
V =

SI Units ] 5000 3 12000 9000

I
C O & to

l2

6000
t > 3000

12

3000

6000

9000

12000

15000

Comparative Method: VITROS 950 System (U/mL)

Comparative Method: VITROS 950 System (U/L)

Method Comparison for CHE DT: Serum


Conventional Units (U/mL) SI Units (U/L) Correlation Range of Range of Slope Coefficient Sample Activity Intercept Sy.x Sample Activity Intercept 1.03 0.998 1.44-12.13 0.13 0.17 1436-12130 131.01

n DT60 II System vs. 950 System 73

Sy.x 172.77

Precision
Precision was evaluated with quality control materials on the VITROS DT60 II System following NCCLS Protocol EP5.15 The data presented are a representation of test performance and are provided as a guideline. Variables such as sample handling and storage, reagent handling and storage, laboratory environment, and system maintenance can affect reproducibility of test results.

Precision for CHE DT: Serum


Mean Activity 3.76 Conventional Units (U/mL) Within Within Day SD* Lab S D " 0.02 0.05 0.04 7.39 0.11 Mean Activity 3760 SI Units (U/L) Within Within Day SD* Lab SD** 23.2 52.8 7391 38.8 105.8 Within Lab CV%** 1.4 1.4 No. Observ. 88 88 No. Days 22 22

System VITROS DT60 II

Within Day precision was determined using two runs/day with two replications. Within Lab precision was determined using a single lot of slides and calibrating weekly.

Pub.

No. C-358

Version 1.0

INSTRUCTIONS FOR USE


References

CHEDT
Cholinesterase

References
1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. MossDW, Henderson AR, Kachman JF. Enzymes. In Textbook of Clinical Chemistry, NWTietz, ed., Philadelphia, PA: WB Saunders;

746-751, 1986.
Trundle D, and Marcial G. Detection of Cholinesterase Inhibition. Annals of Clinical and Laboratory Science. 5:345-352, 1988. NCCLS. Protection of Laboratory Workers from Instrument Biohazards and Infectious Diseases Transmitted by Blood, Body Fluids and Tissue; Approved Guideline. NCCLS Document M29 (OSBIM 1-56238). NCCLS, Wayne, PA 19087; 1997. Calam RR. Specimen Processing Separator Gels: An Update. J Clin Immunoassay. 11:86-90; 1988. Tietz NW (ed). Fundamentals of Clinical Chemistry, ed. 5. Philadelphia: WB Saunders; 387; 2001. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Venipuncture. NCCLS Document H3. Wayne, PA: NCCLS;

1991.
NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Skin Puncture. NCCLS Document H4. Wayne, PA: NCCLS;

1991.
Clinical Laboratory Handbook for Patient Preparation and Specimen Handling. Fascicle VI: Chemistry/Clinical Microscopy. Northfieid, IL: College of American Pathologists; 1992. Method recommended by the "Working Group on Enzymes of the German Society for Clinical Chemistry", European Journal of Clinical Chemistry, Clinical Biochemistry. 30:163-170; 1992. NCCLS. Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition. NCCLS Document C24. Wayne, PA: NCCLS; 1999. NCCLS. Interference Testing in Clinical Chemistry. NCCLS Document EP7. Wayne, PA: NCCLS; 1986. Young DS. Effects of Drugs on Clinical Laboratory Tests, ed. 4. Washington D.C.: AACC Press; 1995. Friedman RB, Young DS. Effects of Disease on Clinical Laboratory Tests. Washington, D.C.: AACC Press; 1990. NCCLS. Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline. NCCLS Document EP9. Wayne, PA: NCCLS; 1995. NCCLS. User Evaluation of Precision Performance with Clinical Chemistry Devices. NCCLS Document EP5. Wayne, PA: NCCLS;

1992.

Glossary of Symbols
The Glossary of Symbols table defines the symbols used on the VITROS Chemistry Products packaging and on the VITROS Chemistry Products Assay Sheets.

Glossary of Symbols
Do Not Reuse Use by or Expiration Date (YYYY-MM-DD) Lot Number Manufacturer's Serial Number Catalog Number or Product Code Attention: See Instructions for Use. Manufacturer Authorized Representative Contains Sufficient for "n" Tests For In Vitro Diagnostic Use Store At or Below Store At or Above Store Between Consult Instructions for Use Fragile, Handle with Care. Keep Dry This end up

Version 1.0

Pub. No. C-358

CHEDT
Cholinesterase

INSTRUCTIONS FOR USE


Revision History
Version 1.0 Description of Technical Changes* New format New organization and sections consistent with IVD Directive Materials Required But Not Provided Sample Dilution - added VITROS 7% BSA; removed reagent-grade water > Known Interfering Substances table - removed ibuprofen; updated values Method Comparison - updated all comparisons and the plot > Precision -updated data > References - added all except 13

Revision History
Date of Revision 2003-10-01

The change bars indicate the position of a technical amendment to the text with respect to the previous version of the document.

When this Instructions For Use is replaced, sign and date below and retain as specified by local regulations or laboratory policies, as appropriate.

Signature

Obsolete Date

C
REP

Ortho-Clinical Diagnostics Mandeville House 62 The Broadway Amersham Buckinghamshire HP7 OHJ England Ortho-Clinical Diagnostics, Inc. 100 Indigo Creek Drive Rochester, NY 14626-5101

'Ortho-Clinical Diagnostics

VITROS is a trademark of Ortho-Clinical Diagnostics, Inc. Ortho-Clinical Diagnostics, Inc., 2003. All rights reserved. Printed in USA.

Pub. No. C-358

Version 1.0

Chemistry

INSTRUCTIONS FOR USE


VITROS Chemistry Products CHOL DT Slides Intended Use

CHOIDT
Cholesterol

For in vitro diagnostic use only. VITROS CHOL DT Slides quantitatively measure cholesterol (CHOL) concentration in serum and plasma.

Summary and Explanation of the Test


Cholesterol is present in tissues and in serum and plasma either as cholesterol or as cholesterol esters bound to proteins. Cholesterol is an essential structural component of cell membranes and the outer layer of plasma lipoproteins and is the precursor of all steroid hormones, including sex and adrenal hormones, bile acids, and vitamin D. Cholesterol measurements are used to evaluate the risk of developing coronary artery occlusion, atherosclerosis, myocardial infarction, and cerebrovascular disease. Coronary atherosclerosis correlates with a high cholesterol level. Cholesterol concentrations are increased in primary hypercholesterolemia; secondary hyperlipoproteinemia, including nephrotic syndrome; primary biliary cirrhosis; hypothyroidism; and in some cases diabetes mellitus. Low cholesterol concentrations may be found in malnutrition, malabsorption, advanced malignancy, and hyperthyroidism. Serum cholesterol concentration depends on many factors, including age and gender.1

Principles of the Procedure


The VITROS CHOL DT Slide method is performed using the VITROS CHOL DT Slide and the VITROS Chemistry Products DT Calibrator Kit on VITROS DT60/DT60 II Chemistry Systems. The VITROS CHOL DT Slide is a multilayered, analytical element coated on a polyester support. The method is based on an enzymatic method similar to that proposed by Allain et al.2 A drop of patient sample is deposited on the slide and is evenly distributed by the spreading layer to the underlying layers. The Triton X-100 (TX100) surfactant in the spreading layer aids in dissociating the cholesterol and cholesterol esters from lipoprotein complexes present in the sample. Hydrolysis of the cholesterol esters to cholesterol is catalyzed by cholesterol ester hydrolase. Free cholesterol is then oxidized in the presence of cholesterol oxidase to form cholestenone and hydrogen peroxide. Finally, hydrogen peroxide oxidizes a leuco dye in the presence of peroxidase to generate a colored dye. The density of dye formed is proportional to the cholesterol concentration present in the sample and is measured by reflectance spectrophotometry.

Reaction Sequence
lipoprotein cholesterol esters + H2O cholesterol + O2 H2O2 + leuco dye cholesterol + cholesterol esters + proteins
cholesterol ester hydrolase

cholesterol + fatty acids cholest-4-en-3-one + H2O2

cholesterol oxidase

peroxidase

- > dye + 2H2O

Test Type and Conditions


Test Type and Conditions for CHOL DT
Test Type Colorimetric VITROS DT60/DT60 II Module DT60/DT60 II Approximate Incubation Time 5 minutes Temperature 37C (98.6F) Wavelength 555 nm Sample Drop Volume 10 uL

Version 1.0

Pub.

No. C-304

CHOL DT
Cholesterol

INSTRUCTIONS FOR USE


Warnings and Precautions

Warnings and Precautions


For in vitro diagnostic use only. Take care when handling materials and samples of human origin. Since no test method can offer complete assurance that infectious agents are absent, consider all clinical specimens, controls, and calibrators potentially infectious. Handle specimens, solid and liquid waste, and test components in accordance with local regulations and NCCLS Guideline M29 or other published biohazard safety guidelines. For specific warnings and precautions for calibrators, quality control materials, and other components, refer to the instructions for use for the appropriate VITROS product, or to other manufacturer's product literature.

Reagents
Slide Diagram

Slide Ingredients

Reactive ingredients per cm

1. Jpper slide mount 2. Spreading layer (BaSO2) 2


> > >
,

Triton X-100 0.8 mg; cholesterol oxidase (Nocardia E.C.1.1.3.6) 0.2 U;


, * " " " " "

Triton X-100 cholesterol ester hydrolase cholesterol oxidase peroxidase leuco dye

- .

cholesterol ester hydrolase (Candida rugosa, E.C.3.1.1.13) 0.7 U; peroxidase (horseradish root, E.C. 1.11.1.7) 5.3 U; and 2-(3,5-dimethoxy4-hydroxyphenyl)-4,5-bis-(4-dimethylaminophenyl) imidazole (leuco dye) 0.2 mg.

'

'

3. leagent layer buffer, pH 6.25 4. Support layer 6. Lower slide mount

Other ingredients
Pigment, binder, buffer, surfactants, stabilizers and cross-linking agent.

Slide Handling
Do not use slides with damaged or incompletely sealed packaging.

Slide Preparation
IMPORTANT: The slide must reach room temperature, 18-28C (64-82F), before the wrapper is opened. Do not use unopened slides that have been at room temperature, 18-28C (64 -82 F) for >48 hours. 1. 2. Remove the unopened slide from the box. Warm the unopened slide for at least 15 minutes. Unopened slides that remain in the sealed wrapper may be returned to refrigerator or freezer storage. 3. Remove the packaging and place the slide into the loading station within 15 minutes after opening.

Slide Storage and Stability


VITROS CHOL DT Slides are stable until the expiration date on the carton when they are stored and handled as specified.

Slide Storage and Stability for CHOL DT


Slides Unopened Storage Condition Room temperature 18-28C (64-82F) Refrigerated 2-8C (36-46F) Frozen <-18C(<0Fl Room temperature 18-28C (64-82F) Stability <48 hours Until expiration date Until expiration date <15 minutes

Opened

Specimen Requirements
G: Handle specimens as biohazardous material.

Specimens Recommended
Serum Plasma: Heparin Certain collection devices have been reported to affect other analytes and tests4 Confirm that your collection devices are compatible with this test.

IMPORTANT:

Pub.

No. C-304

Version 1.0

INSTRUCTIONS FOR USE


Testing Procedure Specimens Not Recommended
Plasma:5 Fluoride oxalate EDTA Sodium citrate

CHO1DT
Cholesterol

Serum and Plasma


Specimen Collection and Preparation
Collect specimens using standard laboratory procedures.6 7 Patient Preparation No special patient preparation is necessary. Special Precautions Centrifuge specimens and remove the serum from the cellular material within 3 hours of collection.8 Storage

Specimen Handling and

Handle specimens as biohazardous material. Handle and store specimens in stoppered containers to avoid contamination and evaporation. Mix samples by gentle inversion and bring to room temperature, 18-28C (64-82F), prior to analysis.

Specimen Storage and Stability for CHOL DT: Serum and Plasma8 Storage Temperature Stability
Room temperature Refrigerated Frozen 18O-28OC(64-82F) 2-8C (36-46F) <-18C (<0F) Not recommended <3 days <3 weeks

Testing Procedure
Materials Provided
. VITROS Chemistry Products CHOL DT Slides

Materials Required But Not Provided


. VITROS Chemistry Products DT Calibrator Kit Quality control materials, such as VITROS Chemistry Products DT Control I & II Isotonic saline Reagent-grade water VITROS DT Pipette

Operating Instructions
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System. Bring all fluids and samples to room temperature, 18-28C (64-82F), prior to analysis. IMPORTANT:

Sample Dilution
If cholesterol concentrations exceed the system's reportable (dynamic) range: 1. Dilute with isotonic saline or reagent-grade water. 2. Reanalyze. 3. Multiply the results by the dilution factor to obtain an estimate of the original sample's cholesterol concentration.

Calibration
Required Calibrators
VITROS Chemistry Products DT Calibrator Kit, bottles 1, 2, and 3

Calibrator Preparation, Handling, and Storage


Refer to the instructions for use for VITROS DT Calibrator Kit.

Calibration Procedure
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Version 1.0

Pub. No. C-304

CHOLDT
Cholesterol When to Calibrate

INSTRUCTIONS FOR USE


Quality Control

Calibrate: When the slide lot number changes. ' When critical system parts are replaced due to service or maintenance. When government regulations require. - For example, in the USA, CLIA regulations require calibration or calibration verification at least once every six months. The VITROS CHOL DT test may also need to be calibrated: If quality control results are consistently outside acceptable range. After certain service procedures have been performed. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Calculations
Reflectance from the slide is measured at 555 nm after the fixed incubation time. Once a calibration has been performed for each slide lot, cholesterol concentration in unknown samples can be determined using the software-resident endpoint colorimetric math model and the response obtained from each unknown test slide.

Validity of a Calibration
Calibration parameters are automatically assessed by the VITROS DT60/DT60 II Chemistry System against a set of quality parameters resident within the analyzer software. Failure to meet any of the pre-defined quality parameters results in a failed calibration. The calibration report should be used in conjunction with quality control results to determine the validity of a calibration.

Reportable (Dynamic) Range Reportable (Dynamic) Range for CHOL DT


Conventional (mg/dL) SI Units (mmol/L) 1.3-8.4

50-325

For out-of-range samples, refer to "Sample Dilution."

Traceability of the Calibration


Values assigned to the VITROS Chemistry Products DT Calibrator Kit for cholesterol are traceable to the Certified NIST (National Institute of Standards and Technology) Reference Material, SRM (Standard Reference Material) 911b. The Ortho-Clinical Diagnostics calibration laboratory uses SRM19 911b to calibrate the Centers for Disease Control (CDC) Modified Abell-Kendall method9 to support cholesterol value assignment for the VITROS DT Calibrator Kit.

Quality Control
Procedure Recommendations
| Handle quality control materials as biohazardous material. Choose control levels that check the clinically relevant range. Analyze quality control materials in the same manner as patient samples, before or during patient sample processing. . To verify system performance, analyze control materials: - After calibration. - According to local regulations or at least once each day that the test is being performed. - After specified service procedures are performed. Refer to the operator's manual for your VITROS DT System. If control results fall outside your acceptable range, investigate the cause before deciding whether to report patient results. For general quality control recommendations, refer to Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition or other published guidelines. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Pub. No. C-304

Version 1.0

INSTRUCTIONS FOR USE


Expected Values and Reporting Units

CHOIDT
Cholesterol

Quality Control Material Selection


IMPORTANT; VITROS DT Control I & II are recommended for use with the VITROS DT60/DT60 II Chemistry System. Evaluate the performance of other commercial control fluids for compatibility with this test before using for quality control.

Control materials other than VITROS DT Controls I & II may show a difference when compared with other Cholesterol methods if they: - Depart from a true human matrix. - Contain high concentrations of preservatives, stabilizers, or other nonphysiological additives. Do not use control materials stabilized with ethylene glycol.

Quality Control Material Preparation and Storage


Refer to the instructions for use for VITROS Chemistry Products DT Control I & II or to other manufacturer's product literature.

Expected Values and Reporting Units


Reference Interval
This reference interval is recommended by NCEP.1

Reference Interval for CHOL DT


Desirable Borderline High High

Conventional Units (mg/dL) <200 200-239 >240

SI Units (mmol/L) <5.2 5.2-6.2 >6.2

Each laboratory should confirm the validity of these intervals for the population it serves.

Reporting Units and Unit Conversion


The VITROS DT60/DT60 II Chemistry System may be programmed to report CHOL DT results in conventional and SI units.

Reporting Units and Unit Conversion for CHOL DT Conventional Units


mg/dL

SI Units mmol/L (mg/dL x 0.02586)

Limitations of the Procedure


Known Interferences
Total protein, 9 g/dL (90 g/L), may elevate results by 4.9%. The VITROS CHOL DT Slide method was screened for interfering substances following NCCLS Protocol EP7." The substances listed in the table, when tested at the concentrations indicated, caused the bias shown.

Known Interfering Substances for CHOL DT


Interferent* Gentisic acid N-acetylcysteine Ethamsylate L-Dopa
*

Interferent Concentration 5 mg/dL 10 mg/dL 3 mg/dL 0.6 mg/dL (0.32 mmol/L) (0.61 mmol/L) (0.114 mmol/L) (0.030 mmol/L)

Cholesterol Concentration Conv. SI (mg/dL) (mmol/L) 210 5.4 210 5.4 210 5.4 210 5.4

Bias Conv. (mg/dL)


-29

-27 -11 -11

SI (mmol/L) -0.8 -0.7 -0.3 -0.3

It is possible that other interfering substances may be encountered. These results are representative; however, your results may differ somewhat due to test-to-test variation. The degree of interference at concentrations other than those listed might not be predictable.

Other Limitations
Certain drugs and clinical conditions are known to alter cholesterol concentration in vivo. For additional information, refer to one of the published summaries.13 u

Version 1.0

Pub. No. C-304

GHOLDT
Cholesterol

INSTRUCTIONS FOR USE


Performance Characteristics

Performance Characteristics
Method Comparison

The plots and table show the results of a comparison of samples analyzed on the VITROS DT60 II Chemistry System with those analyzed using the Modified Abell-Kendall comparative method.9 Testing followed NCCLS Protocol EP9.

Method Comparison for CHOL DT: Serum


Conventional Units

SI Units
10 y =x

350 i
g300

1 I
w

250 H 200

i5

a. >
0 50 100 150 200 250 300 0

100

I
350
Comparative Method: Modified Abell-Kendall (mg/d/L)

10

Comparative Method: Modified Abell-Kendall

Method Comparison for CHOL DT: Serum


Conventional Units (mg/dL) n DT60 II System vs. comparative method 56 Correlation Slope Coefficient 1.00 0.997 Range of Sample Cone. Intercept Sy.x SI Units (mmol/L) Range of Sample Cone. Intercept Sy.x

56-313

0.75

6.11

1.4-8.1

0.02

0.16

Precision
Precision was evaluated with quality control materials on VITROS the DT60 II System following NCCLS Protocol EP5.16 The data presented are a representation of test performance and are provided as a guideline. Variables such as sample handling and storage, reagent handling and storage, laboratory environment, and system maintenance can affect reproducibility of test results.

Precision for CHOL DT: Serum


Mean Cone. 149 235 Conventional Units (mg/dL) Within Within Day SD* Lab S D " 3.2 3.8 3.5 5.0 Mean Cone. 3.8 6.1 SI Units (mmol/L) Within Within Day SD* Lab SD** 0.08 0.10 0.09 0.13 Within Lab CV%** 2.3 2.1 No. Observ. 88 88 No. Days 22 22

System VITROS DT60 II

Within Day precision was determined using two runs/day with two replications. Within Lab precision was determined using a single lot of slides and calibrating weekly.

Pub. No. C-304

Version 1.0

INSTRUCTIONS FOR USE


References

CHOLDT
Cholesterol

References
1. 2. 3. 4. 5. 6. 7. 8. 9. Tietz NW (ed). Fundamentals of Clinical Chemistry, ed. 3. Philadelphia: WB Saunders; 448-468; 1987. Allain CC, et al. Enzymatic Determination of Total Cholesterol in Serum. Clin. Chem. 20:470; 1974. NCCLS. Protection of Laboratory Workers from Instrument Biohazards and Infectious Diseases Transmitted by Blood, Body Fluids and Tissue; Approved Guideline. NCCLS Document M29 (OSBN 1-56238). NCCLS, Wayne, PA 19087; 1997. Calam RR. Specimen Processing Separator Gels: An Update. J Clin Immunoassay. 11:86-90; 1988. NCEP. Recommendations for improving cholesterol measurement. A report from the Laboratory Standardization Panel of the National Cholesterol Education Program. NIH Publication No. 90-2964; 1990. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Venipuncture. NCCLS Document H3. Wayne, PA: NCCLS;

1991.
NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Skin Puncture. NCCLS Document H4. Wayne, PA: NCCLS;

1991.
Burtis CA, Ashwood ER. eds. Textbook of Clinical Chemistry, ed. 3. Philadelphia: WB Saunders; 849; 1999. Duncan IW, Mather A, Cooper GR. The procedure for the proposed cholesterol reference method. Atlanta, GA: Centers for Disease Control; 1982. 10. NCCLS. Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition. NCCLS Document C24. Wayne, PA: NCCLS; 1999. 11. NCEP. Third Report of the National Cholesterol Education Program (NCEP) Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panei III); Executive Summary. NIH Publication No. 01-3670. National Institutes of Health. Bethesda. Maryland: May, 2001. 12. NCCLS. Interference Testing in Clinical Chemistry. NCCLS Document EP7. Wayne, PA: NCCLS; 1986. 13. Young DS. Effects of Drugs on Clinical Laboratory Tests, ed. 4. Washington D C : AACC Press; 1995. 14. Friedman RB, Young DS. Effects of Disease on Clinical Laboratory Tests. Washington, D.C.: AACC Press; 1990. 15. NCCLS. Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline. NCCLS Document EP9. Wayne, PA:

NCCLS; 1995.
16. NCCLS. User Evaluation of Precision Performance with Clinical Chemistry Devices. NCCLS Document EP5. Wayne, PA: NCCLS;

1992.

Glossary of Symbols
The Glossary of Symbols table defines the symbols used on the VITROS Chemistry Products packaging and on the VITROS Chemistry Products Assay Sheets.

Glossary of Symbols
Do Not Reuse Use by or Expiration Date (YYYY-MM-DD) Lot Number Manufacturer's Serial Number Catalog Number or Product Code Attention: See Instructions for Use. j 6c | REP j Manufacturer Authorized Representative Contains Sufficient for "n" Tests For In Vitro Diagnostic Use Store At or Below Store At or Above Store Between Consult Instructions for Use Fragile, Handle with Care. Keep Dry This end up

Version 1.0

Pub. No. C-304

CHOIDT
Cholesterol

INSTRUCTIONS FOR USE


Revision History

Revision History
Date of Revision 2003-03-28 Description of Technical Changes* New format > New organization and sections consistent with IVD Directive Specimens Not Recommended - added the section Serum and Plasma: Special Precautions - "added within 3 hours of collection" > Specimen Handling and Storage - updated all stabilities > Quality Control Material Selection - added statement regarding ethylene glycol Method Comparison - updated comparison values and plots Precision - updated all values > References - added all new references The change bars indicate the position of a technical amendment to the text with respect to the previous version of the document. Version 1.0

When this Instructions For Use is replaced, sign and date below and retain as specified by local regulations or laboratory policies, as appropriate.

Signature

Obsolete Date

C
I E REPj C Ortho-Clinical Diagnostics Mandeville House 62 The Broadway Amersham Buckinghamshire HP7 OHJ England Ortho-Clinical Diagnostics, Inc. 100 Indigo Creek Drive Rochester, NY 14626-5101

' Ortho-Clinical Diagnostics


aAtMSH company

ml

VITROS is a trademark of Ortho-Clinical Diagnostics, Inc. Ortho-Clinical Diagnostics, Inc., 2003. All rights reserved. Printed in USA.

Pub. No. C-304

Version 1.0

[Product*

VITRCD5
Chemistry!

INSTRUCTIONS FOR USE


VITROS Chemistry Products CK DT Slides Intended Use
For in vitro diagnostic use only. VITROS CK DT Slides quantitatively measure creatine kinase (CK) activity in serum and plasma.

CKDT
Creatine Kinase

Summary and Explanation of the Test


Creatine kinase, also referred to as creatine phosphokinase, is a cellular enzyme with a wide tissue distribution. CK is found mainly in skeletal and cardiac muscle. CK's physiological role is associated with ATP generation for contractile or transport systems. Serum CK is almost always increased following acute myocardial infarction or skeletal muscle damage. The enzyme is commonly elevated in myocarditis of any cause, cerebrovascular accidents, rhabdomyolysis, polymyositis, and acute physical exertion. CK is also increased in the muscular dystrophies; in Duchenne's muscular dystrophy, CK elevations of 20-200 times normal are common. Low CK may reflect decreased muscle mass or muscle wasting. Reference values for CK must consider the age, gender, and physical activity of the person. Low serum CK activities are common in the elderly, in the bedridden, and in patients with advanced malignancy.1

Principles of the Procedure


The VITROS CK DT Slide method is performed using the VITROS CK DT Slide and the VITROS Chemistry Products DT Calibrator Kit on VITROS DT60/DT60 II Chemistry Systems. The VITROS CK DT Slide is a multilayered, analytical element coated on a polyester support. A drop of patient sample is deposited on the slide and is evenly distributed by the spreading layer to the underlying layers. This layer also contains W-acetylcysteine (NAC) to activate CK without pretreating the sample. When the sarnple is deposited on the slide, creatine kinase catalyzes the conversion of creatine phosphate and ADP to creatine and ATP. In the presence of glycerol kinase (GK), glycerol is phosphorylated to L-a-glycerophosphate by ATP. Oxidation of L-a-glycerophosphate to dihydroxyacetone phosphate and hydrogen peroxide occurs in the presence of L-a-glycerophosphate oxidase (a-GPO). Finally, leuco dye is oxidized by hydrogen peroxide in the presence of peroxidase to form a dye. Reflection densities are monitored during incubation. The rate of change in reflection density is proportional to enzyme activity.

Reaction Sequence
creatine phosphate + ADP glycerol + ATP L-a-glycerophosphate + O2 H2O2 + leuco dye
peroxidase GK CK NAC, Mg

->

creatine + ATP

L-a-glycerophosphate + ADP
a-GPO

> dihydroxyacetone phosphate + H2O2 dye + 2H2O

Test Type and Conditions


Test Type and Conditions for CK DT
Test Type Rate VITROS DT60/DT60 II Module DTSC/DTSC II Approximate Incubation Time 5 minutes Temperature 37C (98.6F) Wavelength 680 nm Sample Drop Volume 10 uL

Version 1.0

Pub. No. C-342

GKDT
Creatine Kinase

INSTRUCTIONS FOR USE


Warnings and Precautions

Warnings and Precautions


For in vitro diagnostic use only. Take care when handling materials and samples of human origin. Since no test method can offer complete assurance that infectious agents are absent, consider all clinical specimens, controls, and calibrators potentially infectious. Handle specimens, solid and liquid waste, and test components in accordance with local regulations and NCCLS Guideline M292 or other published biohazard safety guidelines. For specific warnings and precautions for calibrators, quality control materials, and other components, refer to the instructions for use for the appropriate VITROS product, or to other manufacturer's product literature.

Reagents
Slide Diagram

Slide Ingredients
" '

12

Jpper slide mount Spreading layer (TiO2) W-acetylcysteine

Reactive ingredients per cm

s2
-"'
... 3

L-alpha-glycerophosphate oxidase (Aerococcus viridans, E.C. 1.1.3.21) 0.4 U; peroxidase (horseradish root, E.C.1.11.1.7) 1.4 U; glycerol kinase (E.coli, E.C.2.7.1.30) 0.5 U; creatine phosphate 170 ug; N-acetylcysteine 54 ug; magnesium acetate 20 ug; glycerol 20 ug; 2-(3,5-dimethoxy-4-hydroxyphenyl)4,5-bis-(4-dimethylaminophenyl) imidazole (leuco dye) 20 ug; and adenosine diphosphate 20 ug.

*-

3. Reagent layer buffer, pH ?.O > adenosine diphosphate > glycerol, magnesium acetate > glycerol kinase, leuco dye peroxidase glycerophosphate oxidase creatine phosphate

Other ingredients
Pigment, binder, buffers, surfactants, inhibitors, stabilizers, cross-linking agent, dye solubilizer, scavenger and chelator.

- - ~S

4.

Support layer

6. -ower slide mouont

Slide Handling
CAUTION: Do not use slides with damaged or incompletely sealed packaging.

Slide Preparation
IMPORTANT: The slide must reach room temperature, 18-2BC (64-82F), before the wrapper is opened. Do not use unopened slides that have been at room temperature, 18 "-28 C (64 -82 F) for >48 hours. 1. 2. Remove the unopened slide from the box. Warm the unopened slide for 15 minutes. Unopened slides that remain in the sealed wrapper may be returned to refrigerator or freezer storage. 3. Remove the packaging and place the slide into the loading station within 15 minutes after opening.

Slide Storage and Stability


VITROS CK DT Slides are stable until the expiration date on the carton when they are stored and handled as specified.

Slide Storage and Stability for CK DT


Slides Unopened

Opened

Storage Condition Room temperature 18-28C (64-82F) Frozen <-18C(<0F) Room temperature 18-28C (64-82F)

Stability <48 hours Until expiration date <15 minutes

Specimen Requirements
v * ::! Handle specimens as biohazardous material.

Specimens Recommended
Serum Plasma:3 Heparin Certain collection devices have been reported to affect other analytes and tests." Confirm that your collection devices are compatible with this test.

IMPORTANT:

Pub. No. C-342

Version 1.0

INSTRUCTIONS FOR USE


Testing Procedure Specimens Not Recommended Do not use grossly hemolyzed specimens.5

CKDT
Creatine Kinase

Serum and Plasma


Specimen Collection and Preparation
Collect specimens using standard laboratory procedures.6 ' Patient Preparation No special patient preparation is necessary. Special Precautions CK is unstable in serum. Centrifuge specimens and remove the serum from the cellular material within 4 hours of collection.8 Specimen Handling and Storage Handle and store specimens in stoppered containers to avoid contamination and evaporation. Mix samples by gentle inversion and bring to room temperature, 18-28C (64-82F), prior to analysis.

Specimen Storage and Stability for CK DT: Serum and Plasma8 Storage Temperature Stability Room temperature 18-28C (64-82F) <4 hours 2-8C (36-46F) Refrigerated <5 days Frozen <-18C(<0F) <1 month

Testing Procedure
Materials Provided
. VITROS Chemistry Products CK DT Slides

Materials Required But Not Provided


. VITROS Chemistry Products DT Calibrator Kit Quality control materials, such as VITROS Chemistry Products DT Control I or VITROS Chemistry Products DT Isoenzyme Control I and VITROS Chemistry Products DT Control II VITROS Chemistry Products 7% BSA VITROS DT Pipette

Operating Instructions
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System. Bring all fluids and samples to room temperature, 18-28C (64-82F), prior to analysis.

I
|

IMPORTANT:

Sample Dilution
If creatine kinase activities exceed the system's reportable (dynamic) range: 1. Dilute the sample with VITROS 7% BSA. 2. Reanalyze. 3. Multiply the results by the dilution factor to obtain an estimate of the original sample's creatine kinase activity. Sample dilution results in higher creatine kinase activities than expected.9

Calibration
Required Calibrators
VITROS Chemistry Products DT Calibrator Kit, bottles 1,2, and 4

Calibrator Preparation, Handling, and Storage


Refer to the instructions for use for VITROS DT Calibrator Kit.

Calibration Procedure
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Version 1.0

Pub. No. C-342

CKDT
Creatine Kinase

INSTRUCTIONS FOR USE


Quality Control
Calibrate: When the slide lot number changes. When critical system parts are replaced due to service or maintenance. When government regulations require. - For example, in the USA, CLIA regulations require calibration or calibration verification at least once every six months. The VITROS CK DT test may also need to be calibrated: If quality control results are consistently outside acceptable range. After certain service procedures have been performed. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

When to Calibrate

Calculations
Based on sequential readings of the slide's reflectance at 680 nm over the defined incubation period, a rate of change in reflectance is determined. This rate is used in the software-resident multi-point rate calibration model to compute enzyme activity. Once a calibration has been performed for each slide lot, creatine kinase activity in unknown samples can be determined from the rate of change in reflectance measured for each unknown test slide

Validity of a Calibration
Calibration parameters are automatically assessed by the VITROS DT60/DT60 II Chemistry System against a set of quality parameters resident within the analyzer software. Failure to meet any of the pre-defined quality parameters results in a failed calibration. The calibration report should be used in conjunction with quality control results to determine the validity of a calibration.

Reportable (Dynamic) Range Reportable (Dynamic) Range for CK DT


Conventional and SI Units (U/L) 20-1600 For out-of-range samples, refer to "Sample Dilution."

Traceability of the Calibration

I
|

Values assigned to the VITROS Chemistry Products DT Calibrator Kit for creatine kinase (CK) are traceable to a modification of the Scandinavian Committee on Enzymes, recommended method 1011 for the determination of creatine kinase at 37C.

Quality Control
Procedure Recommendations
WARNING: Handle quality control materials as biohazardous material. Choose control levels that check the clinically relevant range. Analyze quality control materials in the same manner as patient samples, before or during patient sample processing. To verify system performance, analyze control materials: - After calibration. - According to local regulations or at least once each day that the test is being performed. - After specified service procedures are performed. Refer to the operator's manual for your VITROS DT60/DT60 II System. If control results fall outside your acceptable range, investigate the cause before deciding whether to report patient results. For general quality control recommendations, refer to Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition''2 or other published guidelines. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Pub. No. C-342

Version 1.0

INSTRUCTIONS FOR USE


Expected Values and Reporting Units Quality Control Material Selection
IMPORTANT: VITROS DT Controls are recommended for use with the VITROS DT60/DT60 II Chemistry System. Evaluate the performance of other commercial control fluids for compatibility with this test before using for quality control.

GKDT
Creatine Kinase

Control materials other than VITROS DT Controls may show a difference when compared with other creatine kinase methods if they: - Depart from a true human matrix. - Contain high concentrations of preservatives, stabilizers, or other nonphysiological additives. Do not use control materials stabilized with ethylene glycol.

Quality Control Material Preparation and Storage


Refer to the instructions for use for VITROS Chemistry Products DT Control I & II and VITROS Chemistry Products DT Isoenzyme Control I or to other manufacturer's product literature.

Expected Values and Reporting Units


Reference Interval Reference Interval for CK DT13
Conventional and SI Units (U/L) Females Males 30-135 55-170

The upper limit of the reference interval is reported to be affected by population characteristics such as the degree of physical activity" and race.15 Distributions of CK values from normal, healthy subjects often demonstrate a positive skew,16 leading to variable upper reference limit estimates. Each laboratory should confirm the validity of these intervals for the population it serves.

Reporting Units
The VITROS DT60/DT60 II Chemistry System may be programmed to report creatine kinase results in conventional and SI units.

Reporting Units for CK DT


Conventional and SI Units U/L

Limitations of the Procedure Known Interferences


I Carbon dioxide at a level of 40 mmol/L may cause up to a 30% negative bias in creatine kinase.

Other Limitations
Certain drugs and clinical conditions are known to alter creatine kinase activity in vivo. For additional information, refer to one of the published summaries 17,18

Version 1.0

Pub. No. C-342

CKDT
Creatine Kinase

INSTRUCTIONS FOR USE


Performance Characteristics

Performance Characteristics
Methpd Comparison

The plot and table show the results of a comparison of samples analyzed on the VITROS DT60 II System with those analyzed using the VITROS 950 System. Testing followed NCCLS Protocol EP9.19 Method Comparison for CK DT: Serum Conventional and SI Units
IXOO i y =x

1500 e

1200 900

t
a to o

600
:K)0

0 0 300 600 900 1200 1500 1800


Comparative Method: VITROS 950 System (U/L)

Method Comparison for CK DT: Serum Conventional and SI Units (U/L) n DT60 II System vs. comparative method 53 Slope 1.01 Correlation Coefficient 0.999 Range of Sample Activity 34-1379 Intercept 11.10 Sy.x 17.24

Precision
Precision was evaluated with quality control materials on the VITROS DT60 II System following NCCLS Protocol EP5.20 The data presented are a representation of test performance and are provided as a guideline. Variables such as sample handling and storage, reagent handling and storage, laboratory environment, and system maintenance can affect reproducibility of test results. Precision for System VITROS DT60 II

CKDT: Serum
Conventional and SI Units (U/L) Mean Activity 129 813 Within Day SD* 4.0 13.6 Within Lab SD** Within Lab CV%** 7.9 24.7 6.1 3.0 No Observ. 88 88 No. Days 22 22

Within Day precision was determined using two runs/day with two replications. Within Lab precision was determined using a single lot of slides and calibrating weekly.

Pub. No. C-342

Version 1.0

INSTRUCTIONS FOR USE


References

CKDT
Creatine Kinase

References
1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16. 17. 18. 19. 20. Tietz NW(ed). Fundamentals of Clinical Chemistry, ed. 3. Philadelphia: WB Saunders; 373-377; 1987. NCCLS. Protection of Laboratory Workers from Instrument Biohazards and Infectious Diseases Transmitted by Blood, Body Fluids and Tissue; Approved Guideline. NCCLS Document M29 (OSBN 1-56238). NCCLS, Wayne, PA 19087; 1997. Doumas BT, et al. Differences Between Values for Plasma and Serum in Tests Performed in the Ektachem 700 XR Analyzer, and Evaluation of "Plasma Separator Tubes (PST)." Clin. Chem. 35:1:151-153; 1989. Calam RR. Specimen Processing Separator Gels: An Update. J Clin Immunoassay. 11:86-90; 1988. Tietz NW(ed). Fundamentals of Clinical Chemistry, ed. 3. Philadelphia: WB Saunders; 376-377; 1987. NCCLS. Procedures forthe Collection of Diagnostic Blood Specimens by Venipuncture. NCCLS Document H3. Wayne, PA: NCCLS;

1991.
NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Skin Puncture. NCCLS Document H4. Wayne, PA: NCCLS;

1991.
Clinical Laboratory Handbook for Patient Preparation and Specimen Handling. Fascicle VI: Chemistry/Clinical Microscopy. Northfield, IL: College of American Pathologists; 1992. Tietz NW(ed). Textbook of Clinical Chemistry, ed. 3. Philadelphia: WB Saunders; 662; 1999. Scandinavian Committee on Enzymes: Recommended Method for the Determination of Creatine Kinase in Blood. Scand. J. Clin. Lab. Invest. 39:1-5; 1979. Scandinavian Committee on Enzymes: Recommended Method for the Determination of Creatine Kinase in Blood. Scand. J. Clin. Lab. Invest. 36:711-23; 1976. NCCLS. Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition. NCCLS Document C24. Wayne, PA: NCCLS; 1999. Henry JB. Clinical Diagnosis and Management by Laboratory Methods. Philadelphia: WB Saunders; 2088; 1979. Krahn J. Upper Reference Limit for Creatine Kinase. Clin. Chem. 31(1):158; 1985. Black HR. Quallich H-D, and Garlect CB. Racial Difference in Serum Creatine Kinase Levels. Amer. J. Med. 81:479-487; 1986. Miller WG. Chinchilli HD, Nance WD. Sampling from a Skewed Population Distribution as Exemplified by Estimation of the Creatine Kinase Upper Reference Limit. Clin. Chem. 30(1): 18-23; 1984. Young DS. Effects of Drugs on Clinical Laboratory Tests, ed. 4. Washington D.C.: AACC Press; 1995. Friedman RB. Young DS. Effects of Disease on Clinical Laboratory Tests. Washington, D.C.: AACC Press; 1990. NCCLS. Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline. NCCLS Document EP9. Wayne, PA: NCCLS; 1995. NCCLS. User Evaluation of Precision Performance with Clinical Chemistry Devices. NCCLS Document EP5. Wayne, PA: NCCLS;

1992.

Glossary of Symbols
The Glossary of Symbols table defines the symbols used on the VITROS Chemistry Products packaging and on the VITROS Chemistry Products Assay Sheets.

Glossary of Symbols
Do Not Reuse Use by or Expiration Date (YYYY-MM-DD) Lot Number Manufacturer's Serial Number Catalog Number or Product Code Attention: See Instructions for Use. Manufacturer Authorized Representative Contains Sufficient for "n" Tests For In Vitro Diagnostic Use Store At or Below Store At or Above Store Between Consult Instructions for Use Fragile, Handle with Care. Keep Dry This end up

Version 1.0

Pub. No. C-342

CKDT
Creatine Kinase

INSTRUCTIONS FOR USE


Revision History

Revision History
Date of Revision 2003-10-01 Version 1.0 Description of Technical Changes* New format New organization and sections consistent with IVD Directive Specimens Recommended, Special Precautions - removed the statement regarding EDTA and fluoride oxilate Materials Required But Not Provided and Sample Dilution - added VITROS 7% BSA; removed isotonic saline Sample Dilution - added that dilution may result in higher CK activities than expected Reference Interval - updated the statement regarding the determination of a more specific reference range Known Interferences - added values Method comparison - updated all data and the plot Precision - updated all values References - added all except 1, 9,10,14,15,16,13

The change bars indicate the position of a technical amendment to the text with respect to the previous version of the document.

When this Instructions For Use is replaced, sign and date below and retain as specified by local regulations or laboratory policies, as appropriate.

Signature

Obsolete Date

C
Ortho-Clinical Diagnostics Mandeville House 62 The Broadway Amersham Buckinghamshire HP7 OHJ England Ortho-Clinical Diagnostics, Inc. 100 Indigo Creek Drive Rochester, NY 14626-5101

' Ortho-Clinical Diagnostics


m company

VITROS is a trademark of Ortho-Clinical Diagnostics, Inc. Ortho-Clinical Diagnostics, Inc., 2003. All rights reserved. Printed in USA.

Pub. No. C-342

Version 1.0

Products

VITRCD5
Chemistry

INSTRUCTIONS FOR USE


VITROS Chemistry Products CKMB DT Slides Intended Use
For in vitro diagnostic use only. VITROS CKMB DT Slides quantitatively measure creatine kinase MB (CK-MB) activity in serum.

CKMB DT
Creatine Kinase MB

Summary and Explanation of the Test


The MB isoenzyme of creatine kinase is found primarily in cardiac muscle; however, trace amounts are present in skeletal muscle. CK-MB is elevated in acute myocardial infarction, where the test has its greatest use. CK-MB usually peaks between 12 and 24 hours after myocardial infarction and returns to normal in 48 to 72 hours in an uncomplicated case. CK-MB is also increased in myocarditis, Duchenne's muscular dystrophy, polymyositis, rhabdomyolysis, and other myocardial or myopathic disorders.1

Principles of the Procedure


The VITROS CKMB DT Slide method is performed using the VITROS CKMB DT Slide and the VITROS Chemistry Products DT Isoenzyme Calibrator Kit on VITROS DT60/DT60 II Chemistry Systems. The VITROS CKMB DT Slide is a multilayered, analytical element coated on a polyester support. A drop of patient sample is deposited on the slide and is evenly distributed by the spreading layer to the underlying layers. This layer contains surfactants; N-acetylcysteine (NAC), which activates CK without pretreatment of the sample; and goat antihuman CK-M antibodies, which inhibit CK-MM (muscle) activity and ~50% of the CK-MB (heart) activity. The remaining CK activity represents 50% of the total CK-MB isoenzyme concentration plus any-CK-BB (relatively rare). In the reagent layer, creatine kinase in the sample catalyzes the conversion of creatine phosphate and adenosine diphosphate (ADP) to creatine and adenosine triphosphate (ATP). In the presence of glycerol kinase, glycerol is phosphorylated to L-a-glycerophosphate which is then oxidized to dihydroxyacetone phosphate and H2O2 in the reaction catalyzed by L-a-glycerophosphate oxidase. Finally, leuco dye is oxidized by hydrogen peroxide in the presence of peroxidase to form a dye. The low wavelength light cutoff filter on the slide support minimizes the blank rate effects of incident light during dye development. The rate of change in reflection density is converted to enzyme activity.

Reaction Sequence
CK-MM + CK-MB creatine phosphate + ADP glycerol + ATP L-a-glycerophosphate + O2 H2O2 + leuco dye
anti CK-M antibody

->

CK-M inhibition -> creatine + ATP

creatine kinase-B NAC, glycerol kinase Mg

->

L-a-glycerophosphate + ADP dihydroxyacetone phosphate + H2O2 dye + 2H2O

L-a-glycerophosphate oxidase peroxidase

Test Type and Conditions


Test Type and Conditions for CKMB DT
Test Type Rate VITROS DT60/DT60 II Module DTSC/DTSC II Approximate Incubation Time 5 minutes Temperature 37C (98.6F) Wavelength 680 nm Sample Drop Volume 10 uL

Version 1.0

Pub. No. C-351

CKMBDT
Creatine Kinase MB

INSTRUCTIONS FOR USE


Warnings and Precautions

Warnings and Precautions


For in vitro diagnostic use only. Take care when handling materials and samples of human origin. Since no test method can offer complete assurance that infectious agents are absent, consider all clinical specimens, controls, and calibrators potentially infectious. Handle specimens, solid and liquid waste, and test components in accordance with local regulations and NCCLS Guideline M292 or other published biohazard safety guidelines. For specific warnings and precautions for calibrators, quality control materials, and other components, refer to the instructions for use for the appropriate VITROS product, or to other manufacturer's product literature.

Reagents
Slide Diagram

Slide Ingredients Reactive ingredients per cm


2

" / %

L-a-glycerophosphate oxidase (Aerococcus viridans, E.C.1.1.3.21) 0.39 U; peroxidase (horseradish root, E.C.1.11.1,7) 1.4 U; glycerol kinase (. coli or Cellulomonas sp, E.C.2.7.1.30) 0.45 U; creatine phosphate 0.17 mg; N-acetylcysteine 43 ug; goat anti-human CK-M antibody 0.25 mg; magnesium acetate 68 ug; glycerol 23 ug; 2-(3,5-dimethoxy-4-hydroxyphenyl) -4,5-bis(4-dimethylaminophenyl) imidazole (leuco dye) 20 ug; and adenosine diphosphate 8.4 ug.

7^7- - 4
~"^~--^ 5

"

Other ingredients
Pigment, binder, buffers, surfactants, inhibitors, stabilizers, cross-linking agent, dye solubilizer, filter dyes, scavenger and chelator.

1 . Upper slide m o u n t 2 . S p r e a d i n g layer (TIO2) N-acetylcysteine goat antihuman CK-M antibody Reagent layer adenosine diphosphate Mg acetate leuco dye * glycerol kinase peroxidase L-a-glycerophosphate oxidase creatine phosphate glycerol buffer, pH 7.0 4. Support layer 6. Filter low wavelength light cutoff filter 6. Lower slide mount

Slide Handling
Do not use slides with damaged or incompletely sealed packaging.

Slide Preparation
IMPORTANT: The slide must reach room temperature, 18 - 2 8 C (64 -82 F), before the wrapper is opened. Do not use unopened slides that have been at room temperature, 18 -28 C (64 -82 F) for >48 hours. 1. Remove the unopened slide from the box. 2. Warm the unopened slide for 15 minutes. Unopened slides that remain in the sealed wrapper may be returned to refrigerator or freezer storage. 3. Remove the packaging and place the slide into the loading station within 15 minutes after opening.

Slide Storage and Stability


VITROS CKMB DT Slides are stable until the expiration date on the carton when they are stored and handled as specified.

Slide Storage and Stability for CKMB DT Slides Storage Condition


Unopened Opened Room temperature Frozen Room temperature 18-28C (64-82F) <-18QC (<0F) 18-28C (64-82F)

Stability <48 hours Until expiration date <15 minutes

Pub. No. C-351

Version 1.0

Q VITRtpS

INSTRUCTIONS FOR USE


Specimen Requirements

CKMBDT
Creatine Kinase MB

Specimen Requirements
Handle specimens as biohazardous material.

Specimens Recommended
Serum IMPORTANT; Certain collection devices have been reported to affect other analytes and tests.3 Confirm that your collection devices are compatible with this test.

Specimens Not Recommended


Plasma: EDTA Heparin

Serum
Specimen Collection and Preparation
Collect specimens using standard laboratory procedures.4 5 Patient Preparation No special patient preparation is necessary. Special Precautions Centrifuge specimens and remove the serum from the cellular material within 4 hours of collection.6

Specimen Handling and Storage


WARNING: Handle specimens as biohazardous material.

Handle and store specimens in stoppered containers to avoid contamination and evaporation. Mix samples by gentle inversion and bring to room temperature, 18-28C (64-82F), prior to analysis.

Specimen Storage and Stability for CKMB DT: Serum6 Temperature Storage Room temperature 18-28C(64-82F) Refrigerated 2-8C (36-46F) Frozen <-18C(<0F)

Stability <4 hours <2 days <1 year

Testing Procedure
Materials Provided
VITROS Chemistry Products CKMB DT Slides

Materials Required But Not Provided


VITROS Chemistry Products DT Isoenzyme Calibrator Kit Quality control materials, such as VITROS Chemistry Products DT Isoenzyme Control I VITROS Chemistry Products 7% BSA Reagent-grade water VITROS DT Pipette

Operating Instructions
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System. Bring all fluids and samples to room temperature, 18-28C (64-82F), prior to analysis. IMPORTANT:

Sample Dilution
If Creatine Kinase MB concentrations exceed the system's reportable (dynamic) range: 1. Dilute the sample with VITROS 7% BSA or reagent-grade water. 2. Reanalyze. 3. Multiply the results by the dilution factor to obtain an estimate of the original sample's creatine kinase MB activity.

Version 1.0

Pub. No. C-351

GKMBDT
Creatine Kinase MB

INSTRUCTIONS FOR USE


Calibration

Calibration
Required Calibrators
VITROS Chemistry Products DT Isoenzyme Calibrator Kit

Calibrator Preparation, Handling, and Storage


Refer to the instructions for use for VITROS DT Isoenzyme Calibrator Kit.

Calibration Procedure
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

When to Calibrate
Calibrate: When the slide lot number changes. When critical system parts are replaced due to service or maintenance. When government regulations require. - For example, in the USA, CLIA regulations require calibration or calibration verification at least once every six months. The VITROS CKMB DT test may also need to be calibrated: If quality control results are consistently outside acceptable range. After certain service procedures have been performed. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Calculations

Based on sequential readings of the slide's reflectance at 680 nm over the defined incubation period, a rate of change in

reflectance is determined. This rate is used in the software-resident multi-point rate calibration model to compute enzyme activity. Once a calibration has been performed for each slide lot, creatine kinase activity in unknown samples can be determined from the rate of change in reflectance measured for each unknown test slide.

Validity of a Calibration
Calibration parameters are automatically assessed by the VITROS DT60/DT60 II Chemistry System against a set of quality parameters resident within the analyzer software. Failure to meet any of the pre-defined quality parameters results in a failed calibration. The calibration report should be used in conjunction with quality control results to determine the validity of a calibration.

Reportable (Dynamic) Range Reportable (Dynamic) Range for CKMB DT


Conventional and SI Units (U/L) 1.0-300.0 For out-of-range samples, refer to "Sample Dilution."

Traceability of the Calibration

I
|

Values assigned to the VITROS Chemistry Products DT Isoenzyme Calibrator Kit for creatine kinase MB isoenzyme (CK-MB) are traceable to a CK-M immunoinhibition method with quantitation of the remaining CK-B subunit activity by a modification of the Scandinavian Committee on Enzymes 78 recommended method for the determination of creatine kinase activity at 37C.

Quality Control
Procedure Recommendations
WARNING: Handle quality control materials as blohazardous material. Choose control levels that check the clinically relevant range. Analyze quality control materials in the same manner as patient samples, before or during patient sample processing. To verify system performance, analyze control materials: - After calibration. - According to local regulations or at least once each day that the test is being performed. - After specified service procedures are performed. Refer to the operator's manual for your VITROS DT60/DT60 II System. If control results fall outside your acceptable range, investigate the cause before deciding whether to report patient results.

Pub.

No. C-351

Version 1.0

INSTRUCTIONS FOR USE


Interpretation of Results and Expected Results

CKMRDT
Creatine Kinase MB

For general quality control recommendations, refer to Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition9 or other published guidelines. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Quality Control Material Selection


IIPORWT: VITROS DT Isoenzyme Control I is recommended for use with the VITROS DT60/DT60 II Chemistry System. Evaluate the performance of other commercial control fluids for compatibility with this test before using for quality control.

Control materials other than VITROS DT Isoenzyme Control I may show a difference when compared with other creatine kinase MB methods if they: - Depart from a true human matrix. - Contain high concentrations of preservatives, stabilizers, or other nonphysiological additives. Enzyme activity might also vary with enzyme source, diluent temperature, and activation time during reconstitution. Do not use control materials stabilized with ethylene glycol.

Quality Control Material Preparation and Storage


Refer to the instructions for use for VITROS Chemistry Products DT Isoenzyme Control I or to other manufacturer's product literature.

Interpretation of Results and Expected Results


Interpretation of Results
Elevated CK-MB activities are a sensitive and specific indicator of myocardial infarction (Ml). The diagnosis of myocardial infarction should not be based solely on CK-MB results but should be supported by other clinical and laboratory parameters. Clinical judgment is required to interpret CK-MB results. Judgments based on single determinations are of limited value because the CK-MB peak is of short duration. As with any CK-MB method, if the sample is obtained significantly before or after the peak, the result may be negative. Most clinical studies have recommended that samples be taken every 8 to 12 hours. CK-MB results are usually considered positive when three criteria are met: 1. A significant level of CK-MB activity must be present. A decision value of 16 U/L is recommended.10 Values of less than 16 U/L should be reported as negative for CK-MB and % CK-MB should not be calculated. 2. The CK-MB results should fall between 4% and 25% of the total CK value. If % CK-MB is outside this range, the elevation may have arisen from factors other than myocardial infarction. % CK-MB = CK-MB x 100 CK For example: . CK-MB < 4%. Skeletal muscle contains some CK-MB and significant skeletal muscle damage can result in elevated CK-MB activity. The percentage of CK-MB in muscle is low, however, and CK-MB as a percentage of total CK may remain normal (<4%). CK-MB > 25%. The slide results are actually a measure of the B subunit of CK-MB; therefore, the presence of CK-BB or macro CK type I or type II can cause up to twice the measured CK-MB result that would be expected based on total CK activity. Results greater than 25% may indicate presence of CK-BB or macro CK type I or type II. These results should be confirmed by an alternate method. 3. The rise in CK-MB activity to a peak approximately 18 hours after the infarction and the subsequent fall in activity is characteristic of myocardial infarction. CK variants are relatively stable in circulation and do not show the rise-and-fall pattern. The suggested decision criteria for diagnosis of myocardial infarction have been provided by Dr. T. C. Kwong.10 These criteria are based on a study of 134 patients consecutively admitted to the coronary care unit of the University of Rochester Medical Center in Rochester, New York. Each laboratory should confirm the validity of this protocol for the population it serves. Criteria were chosen to maximize the diagnostic efficiency of the test where: Diagnostic Efficiency (%) = True Positives + True Negatives x 100 Total Patients Decision criteria may vary from laboratory to laboratory depending on age, sex, diet, and racial composition of the population, as well as the prevalence of myocardial infarction. Decision criteria may be adjusted to favor either a positive predictive value or a negative predictive value, depending on the intended use of the assay. The table below shows how different cutoff values for peak CK-MB affects the sensitivity and specificity of the test, compared with an electrophoretic procedure. Use the flowchart in figure 1 as a quick reference guide.

Version 1.0

Pub. No. C-351

VITRCpB El

CKMB DT
Creatine Kinase MB

INSTRUCTIONS FOR USE


Interpretation of Results and Expected Results

An alternative use of the test is to screen samples from patients with possible myocardial infarction and to confirm positive results by an alternative method. A total CK value within the normal range is not a reliable index to exclude analysis of CK-MB.11 Effects on Sensitivity and Specificity of Various Cutoff Values for Peak CK-MB CK-MB Cutoff (U/L) 10 12 14 16 18 20 22 24 Sensitivity (%) 96 96 93 91 85 82 78 78 Specificity (%) 79 89 93 94 95 96 96 98 Efficiency (%) 86 92 93 93 91 90 89 90

Reporting Units and Unit Conversion The VITROS Chemistry System may be programmed to report CK-MB results in conventional, SI, and alternate units. Reporting Units and Unit Conversion for CKMB DT Conventional and SI Units U/L Figure 1. Interpretation of Results: Quick Reference Guide

r alternative diagnoses

Yes
Perform serial CK-MB assays Proper mserpretatjtm af CK-MB results reqyeres serial determinations in all circumstances "Normal" Total CK is not 8 reliable index to exclude CK-MB determ

'
<

Negative tor CK-MB IHtjh U/L M8 liul low ~o MB syo^ftsts skoictiil muscle damage)

""\

--,,

% MB at ieasf 4%?

.^>

-5!

Ntgativft foi CK-Mfl {%, MB greater than 25% suggests macro CK ar CK-BBI

"~"\

<T

%MBI6SSKHS2S%?

J ^ " Resiilts siintiki 1H> confimuHt by an alter native method arid v3luat*)d tJ$i!i(j other cSinicfil and laisorator^'parameters

t
Yes

<C

Rise-and-fall pattern?

~^>-

So

f ^

NegativetorCK-MB (Consistent but elevated CK-MB may be atypical macro or BB1

"\ /

Pub.

No. C-351

Version 1.0

INSTRUCTIONS FOR USE


Limitations of the Procedure

CKMB DT
Creatine Kinase MB

Limitations of the Procedure


Know.n Interferences
CK-BB (present in neonates,12 brain ischemia, cerebellar hematoma, shock,13 and carcinomas14) and macro CK type I and type II falsely elevate CK-MB results. These can be differentiated from true CK-MB by the lack of the characteristic CK-MB time pattern. Refer to "Interpretation of Results." Total CK activity greater than 1000 U/L may result in falsely elevated CK-MB results. Samples with TCK >1000 U/L should be diluted prior to analysis. The VITROS CKMB DT Slide method was screened for interfering substances following NCCLS Protocol EP7.16 The substances listed in the table, when tested at the concentrations indicated, caused the bias shown.

Known Interfering Substances for CKMB DT


Interferent* Ascorbic acid Diatrizoate sodium (Hypaque) Dipyrone Interferent Concentration 3mg/dL 835mg/dL 6mg/dL (170 umol/L) (13.1 mmol/L) (180 Mmol/L) Creatine Kinase Activity Conv./SI Units (U/L) 40 40 48 Average Bias Conv./SI Units (U/L) -6 -12 -4.3

It is possible that other interfering substances may be encountered. These results are representative; however, your results may differ somewhat due to test-to-test variation. The degree of interference at concentrations other than those listed might not be predictable.

Other Limitations
Certain drugs and clinical conditions are known to alter creatine kinase MB activity in vivo. For additional information, refer to one of the published summaries.16 17 18

Performance Characteristics
Method Comparison
The plot and table show the results of a comparison of samples analyzed on the VITROS DT60 II System with those analyzed using the Immunoinhibition comparative method with quantitation of the remaining CK-B subunit activity by a modification of the Scandinavian Committee on Enzymes.7 8 Testing followed NCCLS Protocol EP9.19

Method Comparison for CKMB DT: Serum


Conventional and SI Units
y =x

."mi 25ii 200


ISO too

50 0 10 100 150 300 250 300 350


Comparative Method: Immunoinhibition (U/L)

Version 1.0

Pub. No. C-351

CKMBDT
Creatine Kinase MB Method Comparison for CKMB DT: Serum

INSTRUCTIONS FOR USE


References

Conventional and SI Units (U/L) n DT60 II System vs. comparative method 77 Correlation Range of Slope Coefficient Sample Activity Intercept 1.00 0.999 Sy.x

2-292

0.41

4.32

Precision
Precision was evaluated with quality control materials on the VITROS DT60 II System following NCCLS Protocol EP5.20 The data presented are a representation of test performance and are provided as a guideline. Variables such as sample handling and storage, reagent handling and storage, laboratory environment, and system maintenance can affect reproducibility of test results.

Precision for CKMB DT: Serum


Conventional and SI Units (U/L) System VITROS DT60 II Mean Activity 11 29 Within Day SD* 0.3 0.6 Within Lab SD** 0.6 1.0 Within Lab CV%** 5.5 3.5 No. Observ. 88 88 No. Days 22 22

Within Day precision was determined using two runs/day with two replications. Within Lab precision was determined using a single lot of slides and calibrating weekly.

References
Tietz NW (ed). Fundamentals of Clinical Chemistry, ed. 3. Philadelphia: WB Saunders; 383-384; 1987. NCCLS. Protection of Laboratory Workers from Instrument Biohazards and Infectious Diseases Transmitted by Blood, Body Fluids and Tissue; Approved Guideline. NCCLS Document M29 (OSBN 1-56238). NCCLS, Wayne, PA 19087; 1997. 3. Calam RR. Specimen Processing Separator Gels: An Update. J Clin Immunoassay. 11:86-90; 1988. 4. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Venipuncture. NCCLS Document H3. Wayne, PA: NCCLS; 1991. 5. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Skin Puncture. NCCLS Document H4. Wayne, PA: NCCLS; 1991. 6. Clinical Laboratory Handbook for Patient Preparation and Specimen Handling. Fascicle VI: Chemistry/Clinical Microscopy. Northfield, IL: College of American Pathologists; 1992. 7. Scandinavian Committee on Enzymes: Recommended Method for the Determination of Creatine Kinase in Blood. Scand. J. Clin. Lab. Invest. 36:711-723; 1976. 8. Scandinavian Committee on Enzymes: Recommended Method for the Determination of Creatine Kinase in Blood. Scand. J. Clin. Lab. Invest. 39:1-5; 1979. 9. NCCLS. Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition. NCCLS Document C24. Wayne, PA: NCCLS; 1999. 10. Kwong TC. Studies conducted at the University of Rochester Medical Center, Rochester, New York; 1986. 11. Yusuf S, et al. Significance of Elevated MB Isoenzyme with Normal Creatine Kinase in Acute Myocardial Infarction. Am. J. Cardiol. 59:245; 1989. 12. Jedukin R, et al. Creatine Kinase Isoenzymes in Serum from Cord Blood and the Blood of Healthy Full-Term Infants during the First Three Postnatal Days. Clin. Chem. 28:2; 1982. 13. Gerhardt W, et al. Creatine Kinase and Creatine Kinase B-Subunit Activity in Serum Cases of Suspected Myocardial Infarction. Clin. Chem. 28:2; 1982. 14. Stein W, et al. Macro Creatine Kinase Type 2: Results of a Prospective Study in Hospitalized Patients. Clin. Chem. 31:12; 1985. 15. NCCLS. Interference Testing in Clinical Chemistry. NCCLS Document EP7. Wayne, PA: NCCLS; 1986. 16. Young DS. Effects of Drugs on Clinical Laboratory Tests, ed. 4. Washington D.C.: AACC Press; 1995. 17. Friedman RB, Young DS. Effects of Disease on Clinical Laboratory Tests. Washington, D.C.: AACC Press; 1990. 18. Tryding N, Tufvesson C, Sonntag O (eds). Drug Effects in Clinical Chemistry, ed. 7. Stockholm: The National Corporation of Swedish Pharmacies, Pharmasoft AB, Swedish Society for Clinical Chemistry; 1996. 19. NCCLS. Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline. NCCLS Document EP9. Wayne, PA: NCCLS; 1995. 20. NCCLS. User Evaluation of Precision Performance with Clinical Chemistry Devices. NCCLS Document EP5. Wayne, PA: NCCLS; 1992. 1. 2.

Pub. No. C-351

Version 1.0

INSTRUCTIONS FOR USE


Glossary of Symbols

CKMBDT
Creatine Kinase MB

Glossary of Symbols
The Glossary of Symbols table defines the symbols used on the VITROS Chemistry Products packaging and on the VITROS Chemistry Products Assay Sheets.

Glossary of Symbols
Do Not Reuse Use by or Expiration Date (YYYY-MM-DD) Lot Number A M Manufacturer's Serial Manufacturer Authorized Representative Contains Sufficient for "n" Tests For In Vitro Diagnostic Use Store Between Consult Instructions for Use Fragile, Handle with Care. Keep Dry This end up

OlN

Number
Catalog Number or Product Code Attention: See Instructions for Use.

Store At or Below Store At or Above

Revision History
Date of Revision 2003-04-30 Version . 10 Description of Technical Changes* New format New organization and sections consistent with IVD Directive Specimens Not Recommended - removed fluoride oxalate Specimen Storage and Stability - updated stability values Materials Required But Not Provided and Sample Dilution - added VITROS 7% BSA; deleted isotonic saline Limitations of the Procedure - updated values for Dipyrone in the Known Interfering Substances table Method Comparison - updated all data and the plot Precision - updated all data References - added 1 , 2 , 3 , 4 , 5 , 6 , 9 , 15,18, 19,20

The change bars indicate the position of a technical amendment to the text with respect to the previous version of the document.

When this Instructions For Use is replaced, sign and date below and retain as specified by local regulations or laboratory policies, as appropriate.

Signature

Obsolete Date

Version 1.0

Pub. No. C-351

CKMBDT
Creatine Kinase MB

INSTRUCTIONS FOR USE


Revision History

C
EC

Ortho-Clinical Diagnostics Mandeville House 62 The Broadway Amersham Buckinghamshire HP7 OHJ England Ortho-Clinical Diagnostics, Inc. 100 Indigo Creek Drive Rochester, NY 14626-5101

'Ortho-Clinical Diagnostics
8 ^otuwoH^efwHum company

VITROS is a trademark of Ortho-Clinical Diagnostics, Inc. Ortho-Clinical Diagnostics, Inc., 2003. All rights reserved. Printed in USA.

10

Pub.

No. C-351

Version 1.0

(Products;

VITRIIIS
Chemistry!

INSTRUCTIONS FOR USE


VITROS Chemistry Products CI" DT Slides Intended Use
For in vitro diagnostic use only. VITROS Cl" DT Slides quantitatively measure chloride (Cl) concentration in serum and plasma.

Cl DT
Chloride

Summary and Explanation of the Test


Chloride is the major anion in the extracellular water space; its physiological significance is in maintaining proper body water distribution, osmotic pressure, and normal anion-cation balance in the extracellular fluid compartment. Chloride is increased in dehydration, in renal tubular acidosis (hyperchloremia metabolic acidosis), and in excessive infusion of isotonic saline. Chloride is decreased in overhydration, chronic respiratory acidosis, salt-losing nephritis, metabolic alkalosis, and congestive heart failure.1

Principles of the Procedure


The VITROS Cl" DT Slide method is performed using the VITROS Cl" DT Slide and the VITROS Chemistry Products DT Calibrator Kit on VITROS DT60/DT60 II Chemistry Systems. The VITROS Cl" DT Slide is a multilayered, analytical element coated on a polyester support that uses direct potentiometry2 for measurement of chloride ions. The slide consists of two ion-selective electrodes, each containing a protective layer, a silver layer and a silver chloride layer coated on a polyester support. The protective layer inhibits interference from normal levels of bromide and uric acid. A drop of patient sample and a drop of VITROS DT Reference Fluid on separate halves of the slide results in migration of both fluids toward the center of the paper bridge. A stable liquid junction is formed connecting the reference electrode to the sample indicator electrode. Each electrode produces an electrical potential in response to the activity of chloride ions applied to it. The potential difference poised between the two electrodes is proportional to the chloride concentration in the sample.

Test Type and Conditions


Test Type and Conditions for Cl DT
Test Type Potentiometric VITROS DT60/DT60 II Module DTE/DTE II Approximate Incubation Time 180 seconds Temperature 25C (77F) Drop Volume Sample: Reference Fluid: 10 uL 10 ML

Warnings and Precautions


For in vitro diagnostic use only. Take care when handling materials and samples of human origin. Since no test method can offer complete assurance that infectious agents are absent, consider all clinical specimens, controls, and calibrators potentially infectious. Handle specimens, solid and liquid waste, and test components in accordance with local regulations and NCCLS Guideline M293 or other published biohazard safety guidelines. For specific warnings and precautions for calibrators, quality control materials, and other components, refer to the instructions for use for the appropriate VITROS product, or to other manufacturer's product literature.

Version 1.0

Pub. No. C-309

Cl DT
Chloride

INSTRUCTIONS FOR USE


Reagents

Reagents
Slide Diagram

Slide Ingredients
Reactive ingredients per cm2
Silver 0.4 mg; and silver chloride 0.2 mg.

Other ingredients
Polymer, plasticizer, surfactant and nickel.

Upper frame Paper bridge Protective layer Sliver, silver chloride layer 6. Support layer 6. Lower frame

1. 2. 3. 4.

Slide Handling
CAUTION: Do not use slides with damaged or incompletely sealed packaging.

Slide Preparation
IMPORTANT: The slide must reach room temperature, 18 -28 C (64 -82 F), before the wrapper is opened. Do not use unopened slides that have been at room temperature, 18 -28 C (64-82F) for >48 hours. 1. 2. Remove the unopened slide from the box. Warm the unopened slide for 15 minutes. Unopened slides that remain in the sealed wrapper may be returned to refrigerator or freezer storage. 3. Remove the packaging and place the slide into the loading station within 15 minutes after opening.

Slide Storage and Stability


VITROS CT DT Slides are stable until the expiration date on the carton when they are stored and handled as specified.

Slide Storage and Stability for Cl DT


Slides Unopened Storage Condition Room temperature 18-28C (64-82F) Refrigerated 2-8C (36-46F) Frozen <-18C (<0F) Room temperature 18O-28C (64-82F) Stability <48 hours Until expiration date Until expiration date <15 minutes

Opened

Specimen Requirements
WARNING: Handle specimens as biohazardous material.

Specimens Recommended
Serum Plasma: Heparin Certain collection devices have been reported to affect other analytes and tests.4 Confirm that your collection devices are compatible with this test.

IMPORTANT:

Specimen Collection and Preparation


Collect specimens using standard laboratory procedures.5 6 Patient Preparation No special patient preparation is necessary. Special Precautions Do not draw specimen from an arm receiving an intravenous transfusion. Fibrin clots may cause incomplete sampling of the specimen.7 - Allow specimens to clot completely in order to prevent fibrin clots. - Inspect plasma specimens for the presence of fibrin clots. e Centrifuge specimens and remove the serum from the cellular material within 4 hours of collection.'

Pub.

No. C-309

Version 1.0

INSTRUCTIONS FOR USE


Testing Procedure Specimen Handling and Storage Handle specimens as biohazardous material. Handle and store specimens in stoppered containers to avoid contamination and evaporation. Mix samples by gentle inversion and bring to room temperature, 18-28C (64-82F), prior to analysis. Specimen Storage and Stability for Cl DT; Serum and Plasma8 Temperature Stability Storage 18-28C(64-82F) Room temperature <7 days Refrigerated 2-8C (36-46F) <4 weeks Indefinite Frozen <-18C (<0F)

Cl DT
Chloride

Testing Procedure
Materials Provided
VITROS Chemistry Products Cl" DT Slides

Materials Required But Not Provided


VITROS Chemistry Products DT Calibrator Kit Quality control materials, such as VITROS Chemistry Products DT Control I & II VITROS Chemistry Products DT Reference Fluid VITROS DTE Dual Sample Cups VITROS DTE Pipette

Operating Instructions
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System. Bring all fluids and samples to room temperature, 18-28C (64-82F), prior to analysis.

MPORTANT:

Sample Dilution
Chloride concentrations outside the reportable (dynamic) range are not expected. Diluted samples should not be analyzed with VITROS Cl" DT Slides because dilution changes both the concentration of solids in plasma water and the ionic strength of the sample.

Calibration
Required Calibrators
VITROS Chemistry Products DT Calibrator Kit, bottles 1 and 2

Calibrator Preparation, Handling, and Storage


Refer to the instructions for use for VITROS DT Calibrator Kit.

Calibration Procedure
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System. NOTE: Calibrate choloride in duplicate by running each bottle twice.

When to Calibrate
Calibrate: When the slide lot number changes. When critical system parts are replaced due to service or maintenance. When government regulations require. - For example, in the USA, CLIA regulations require calibration or calibration verification at least once every six months. When the VITROS DT Reference Fluid lot number changes. The VITROS Cl" DT test may also need to be calibrated: If quality control results are consistently outside acceptable range. After certain service procedures have been performed. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Version 1.0

Pub. No. C-309

Cl DT
Chloride

INSTRUCTIONS FOR USE


Quality Control
The VITROS DT60/DT60 II Chemistry System measures the potential difference in millivolts between the two electrodes of a potentiometric slideone in contact with the sample to be analyzed and the other in contact with the electrolyte reference fluid. A linear relationship exists between the measured potential difference observed on the slide and the logarithm of chloride concentration, i.e. the Nernst equation for ion-selective electrodes. Once the calibration has been established for each slide lot, unknown chloride concentrations for a given sample can be determined using the software-resident math model and the measured potential difference.

Calculations

Validity of a Calibration
Calibration parameters are automatically assessed by the VITROS DT60/DT60II Chemistry System against a set of quality parameters resident within the analyzer software. Failure to meet any of the pre-defined quality parameters results in a failed calibration. The calibration report should be used in conjunction with quality control results to determine the validity of a calibration.

Reportable (Dynamic) Range Reportable (Dynamic) Range for CI" DT


Conventional and SI Units (mmol/L) 65-140 For out-of-range samples, refer to "Sample Dilution."

Traceability of the Calibration


I I Values assigned to the VITROS Chemistry Products DT Calibrator Kit for chloride are traceable to the Certified NIST (National Institute of Standards and Technology) Reference Material, SRM (Standard Reference Material) 919a. The Ortho-Clinical Diagnostics calibration laboratory uses SRM 919a to calibrate the coulometric-amperometric titration method9'10 to support chloride value assignment for the VITROS DT Calibrator Kit.

Quality Control
Procedure Recommendations
| ."">,. Handle quality control materials as biohazardous material. Choose control levels that check the clinically relevant range. Analyze quality control materials in the same manner as patient samples, before or during patient sample processing. To verify system performance, analyze control materials: - After calibration. - According to local regulations or at least once each day that the test is being performed. - After specified service procedures are performed. Refer to the operator's manual for your VITROS DT60/DT60 II System. If control results fall outside your acceptable range, investigate the cause before deciding whether to report patient results. For general quality control recommendations, refer to Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition'1'1 or other published guidelines. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Quality Control Material Selection


I IMPORTANT; VITROS DT Control I & II are recommended for use with the VITROS DT60/DT60II Chemistry System. Evaluate the performance of other commercial control fluids for compatibility with this test before using for quality control.

Control materials other than VITROS DT Controls I & II may show a difference when compared with other chloride methods if they: - Depart from a true human matrix. - Contain high concentrations of preservatives, stabilizers, or other nonphysiological additives. Do not use control materials stabilized with ethylene glycol.

Quality Control Material Preparation and Storage


Refer to the instructions for use for VITROS Chemistry Products DT Control I & II or to other manufacturer's product literature.

Pub. No. C-309

Version 1.0

INSTRUCTIONS FOR USE


Expected Values and Reporting Units

CIDT
Chloride

Expected Values and Reporting Units


Reference Interval
This reference interval is the central 95% of results from an internal study of 60 apparently healthy individuals from a working population. No significant differences between results from the male and female populations were observed.

Reference Interval for CI" DT


Conventional and SI Units (mmol/L) Adult 98-107

Each laboratory should confirm the validity of these intervals for the population it serves.

Reporting Units and Unit Conversion


The VITROS DT60/DT60 II Chemistry System may be programmed to report chloride results in conventional and SI units.

Reporting Units for CI" DT Conventional and SI Units


mmol/L

Limitations of the Procedure


Known Interferences
Bromide and iodide from therapeutic drugs and ointments may cause a bias of approximately 3 mmol/L and 4 mmol/L, respectively, for each mmol of halide. Normal physiological levels of bromide and iodide do not interfere.

Other Limitations
Certain drugs and clinical conditions are known to alter chloride concentration in vivo. For additional information, refer to one of the published summaries.1213

Performance Characteristics
Method Comparison

The plot and table show the results of a comparison of samples analyzed on the VITROS DT60 II System with those analyzed using the VITROS 950 System. Testing followed NCCLS Protocol EP9.14

Method Comparison for CI DT: Serum Conventional and SI Units


175
y=x

1 50 125
100

75

SO

75

100

125

150

175

Comparative Method: VITROS 950 (mmol/L)

Version 1.0

Pub. No. C-309

Cl DT
Chloride Method Comparison for Cl' DT: Serum

INSTRUCTIONS FOR USE


References

Conventional and SI Units (mmol/L) n DT60 II System vs. comparative method 51 Slope 0.96 Correlation Coefficient 0.995 Range of Sample Activity Intercept 5.67 Sy.x 1.24

75-132

Precision
Precision was evaluated with quality control materials on the VITROS DT60 II System following NCCLS Protocol EP5.15 The data presented are a representation of test performance and are provided as a guideline. Variables such as sample handling and storage, reagent handling and storage, laboratory environment, and system maintenance can affect reproducibility of test results.

Precision for Cl DT: Serum


Conventional and SI Units (mmol/L) System VITROS DT60 II Mean Cone. 78 100 Within Day SD* 0.2 0.4 Within Lab SD** 0.8 1.2 Within Lab CV%** 1.1 1.2 No. Observ. 88 88 No. Days 22 22

Within Day precision was determined using two runs/day with two replications. Within Lab precision was determined using a single lot of slides and calibrating weekly.

References
1. 2. 3. Tietz NW(ed). Fundamentals of Clinical Chemistry, ed. 3. Philadelphia: WB Saunders; 620-621; 1987. Siggard-Anderson O. Electrochemistry, in Tietz NW (ed). Textbook of Clinical Chemistry. Philadelphia: WB Saunders; 110-125; 1986. NCCLS. Protection of Laboratory Workers from Instrument Biohazards and Infectious Diseases Transmitted by Blood, Body Fluids and Tissue; Approved Guideline. NCCLS Document M29 (OSBN 1-56238). NCCLS, Wayne, PA 19087; 1997. 4. Calam RR. Specimen Processing Separator Gels: An Update. J Clin Immunoassay. 11:86-90; 1988. 5. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Venipuncture. NCCLS Document H3. Wayne, PA: NCCLS; 1991. 6. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Skin Puncture. NCCLS Document H4. Wayne, PA: NCCLS; 1991. 7. Slockbower JM, Blumenfeld TA (ed.). Collection and Handling of Laboratory Specimens. Philadelphia: Lippincott Co; 201; 1983. 8. Clinical Laboratory Handbook for Patient Preparation and Specimen Handling. Fascicle VI: Chemistry/Clinical Microscopy. Northfield, IL: College of American Pathologists; 1992. 9. Dietz AA, Bond EE, Chloride, Coulomeric-amperometric Methods, in: Faulkner WR, Meites S, eds. Selected Methods of Clinical Chemistry, Washington: American Association for Clinical Chemistry. 9:149-152.; 1982. 10. Velapoldi RA, Paule RC, Schaffer R, Mandel TJ, Gramlich JW. Standard reference materials: A reference method for the determination of chloride in serum. National Institute of Standards and Technology Special Publication 260-67. Washington, D.C.; 1979. 11. NCCLS. Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition. NCCLS Document C24. Wayne, PA: NCCLS; 1999. 12. Young DS. Effects of Drugs on Clinical Laboratory Tests, ed. 4. Washington D.C.: AACC Press; 1995. 13. Friedman RB, Young DS. Effects of Disease on Clinical Laboratory Tests. Washington, D.C.: AACC Press; 1990. 14. NCCLS. Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline. NCCLS Document EP9. Wayne, PA: NCCLS; 1995. 15. NCCLS. User Evaluation of Precision Performance with Clinical Chemistry Devices. NCCLS Document EP5. Wayne, PA: NCCLS; 1992.

Pub. No. C-309

Version 1.0

INSTRUCTIONS FOR USE


Glossary of Symbols

CIDT
Chloride

Glossary of Symbols
The Glossary of Symbols table defines the symbols used on the VITROS Chemistry Products packaging and on the VITROS Chemistry Products Assay Sheets.

Glossary of Symbols
Do Not Reuse Manufacturer Authorized Representative Contains Sufficient for "n" Tests For In Vitro Diagnostic Use Store At or Below

2
SN
"RIFI

Use by or Expiration Date (YYYY-MM-DD) Lot Number Manufacturer's Serial Number Catalog Number or Product Code Attention: See Instructions for Use.

X I

Store Between Consult Instructions for Use Fragile, Handle with Care. Keep Dry This end up

A
Revision History
Date of Revision 2003-10-01

Store At or Above

Version 1.0

Description of Technical Changes* New format New organization and sections consistent with IVD Directive Specimens Recommended - updated wording Specimen Storage and Stability - updated stability values Quality Control Material Selection - added the statement regarding ethylene glycol Method Comparison - updated all data and the plot Precision - updated all data References - added all

The change bars indicate the position of a technical amendment to the text with respect to the previous version of the document.

When this Instructions For Use is replaced, sign and date below and retain as specified by local regulations or laboratory policies, as appropriate.

Signature

Obsolete Date

Version 1.0

Pub. No. C-309

VITRIJ35 0

CIDT
Chloride

INSTRUCTIONS FOR USE


Revision History

C
EC

I REP

Ortho-Clinical Diagnostics Mandeville House 62 The Broadway Amersham Buckinghamshire HP7 OHJ England Ortho-Clinical Diagnostics, Inc. 100 Indigo Creek Drive Rochester, NY 14626-5101

'Ortho-Clinical Diagnostics
^e&tMroH company

VITROS is a trademark of Ortho-Clinical Diagnostics, Inc. Ortho-Clinical Diagnostics, Inc., 2003. All rights reserved. Printed in USA.

Pub. No. C-309

Version 1.0

Products

VITRCD5
Chemistry!

INSTRUCTIONS FOR USE


VITROS Chemistry Products CO2 DT Slides Intended Use

CO2DT
Carbon Dioxide

For in vitro diagnostic use only. VITROS CO2 DT Slides quantitatively measure carbon dioxide (CO2) concentration in serum and plasma.

Summary and Explanation of the Test


Total carbon dioxide measurements are used together with other clinical and laboratory information (pH, pCO2) for the evaluation of acid-base status.

Principles of the Procedure


The VITROS CO2 DT Slide method is performed using the VITROS CO2 DT Slides and the VITROS Chemistry Products DT Calibrator Kit on VITROS DT60/DT60 II Chemistry Systems. The VITROS CO2 DT Slide is a multilayered, analytical element coated on a polyester support that uses direct potentiometry for measurement of ionic carbon dioxide. The slide consists of two ion-selective electrodes, each containing a buffer layer, an ion selective membrane layer, a reference layer, and a silver and a silver chloride layer coated on a polyester support. The buffer layer adjusts the sample pH to 8.4 and maintains CO2, HCO3", and CO3"2 in proper equilibrium. The ion-selective membrane layer contains an ion exchanger trioctylpropylammonium chloride (TOPA Cl) and a CO3'2 ionophore decyltrifluoroacetophenone(DTFA). A drop of patient sample and a drop of VITROS DT Reference Fluid on separate halves of the slide results in migration of both fluids toward the center of the paper bridge. A stable liquid junction is formed connecting the reference electrode to the sample indicator electrode. Each electrode produces an electrical potential in response to the concentration of carbon dioxide applied to it. The potential difference poised between the two electrodes is proportional to the carbon dioxide concentration in the sample.

Test Type and Conditions


Test Type and Conditions for CO2 DT
Test Type Potentiometric VITROS DT60/DT60 II Module DTE/DTE II Approximate Incubation Time 3 minutes Temperature 25C (77F) Drop Volume Sample: Reference 10 ML Fluid: 10 |JL

Warnings and Precautions


For in vitro diagnostic use only. Take care when handling materials and samples of human origin. Since no test method can offer complete assurance that infectious agents are absent, consider all clinical specimens, controls, and calibrators potentially infectious. Handle specimens, solid and liquid waste, and test components in accordance with local regulations and NCCLS Guideline M291 or other published biohazard safety guidelines. For specific warnings and precautions for calibrators, quality control materials, and other components, refer to the instructions for use for the appropriate VITROS product, or to other manufacturer's product literature.

Version 1.0

Pub.

No. C-308

CO2DT
Carbon Dioxide

INSTRUCTIONS FOR USE


Reagents

Reagents
Slide Diagram

Slide Ingredients
Reactive ingredients per cm
2

Silver 0.4 mg; silver chloride 0.2 mg; sodium chloride 0.2 mg; potassium chloride 63 ng; trioctylpropylammonium chloride 0.3 mg; and decyltrifluoroacetophenone 0.8 mg.
* - .

Other ingredients
Binders, plasticizers, surfactants, stabilizer, buffers and nickel.
T

b G

1. Upper frame 2. Paper Bridge 3. Ion-selective membrane TOPA Cl DTFA 4. Reference layer KCI NaCI 6. Silver, sliver chloride layer 6. Support layer 7. Lower frame

Slide Handling
Do not use slides with damaged or incompletely sealed packaging.

Slide Preparation
The slide must reach room temperature, 18-28C (64-82f), opened. before the wrapper is

Do not use unopened slides that have been at room temperature, 18 -28 C (64-82F) for >48 hours. 1. 2. Remove the unopened slide from the box. Warm the unopened slide for 15 minutes. Unopened slides that remain in the sealed wrapper may be returned to refrigerator or freezer storage. 3. Remove the packaging and place the slide into the loading station within 15 minutes after opening.

Slide Storage and Stability


VITROS CO2 DT Slides are stable until the expiration date on the carton when they are stored and handled as specified.

Slide Storage and Stability for CO2 DT


Slides Unopened Storage Condition 18-28C (64-82F) Room temperature Refrigerated 2-8 c C (36-46F) Frozen <-18C (<0F) 18O-28C(64-82F) Room temperature Stability <48 hours Until expiration date Until expiration date <15 minutes

Opened

Specimen Requirements
WARNING: Handle specimens as biohazardous material.

Specimens Recommended
Serum Plasma:2 Heparin Certain collection devices have been reported to affect other analytes and tests3 Confirm that your collection devices are compatible with this test.

IMPORTANT:

Serum and Plasma


Specimen Collection and Preparation
Collect specimens using standard laboratory procedures.4 6 NOTE: For details on minimum fill volume requirements, refer to the operator's manual for your VITROS Chemistry System.

Patient Preparation No special patient preparation is necessary.

Pub. No. C-308

Version 1.0

VITR^S

INSTRUCTIONS FOR USE


Testing Procedure

CO2DT
Carbon Dioxide

Special Precautions Do not draw specimen from an arm receiving an intravenous transfusion. Do not use specimens from patients receiving radiographic contrast agents containing diatriazoate sodium. Refer to "Limitations of the Procedure." Every effort should be made to fill vacuum tubes completely when collecting blood because a decrease of up to 3 mmol/L can be observed with partially filled tubes.6 Fibrin clots may cause incomplete sampling of the specimen.7 - Allow specimens to clot completely in order to prevent fibrin clots. - Inspect plasma specimens for fibrin clots. Centrifuge specimens and remove the serum from the cellular material within 4 hours of collection.8 Specimen Handling and Storage

Handle specimens as biohazardous material. Handle and store specimens in stoppered containers to avoid contamination and evaporation. Mix samples by gentle inversion and bring to room temperature, 18-28C (64-82F), prior to analysis.

Specimen Storage and Stability for CO2 DT: Serum and Plasma8
Storage Room temperature, tightly capped Refrigerated Frozen Temperature 18 0 -28 0 C(64 0 -82 0 F) 2-8C (36-46F) <-18C(<0F) Stability <24 hours <3 days <1 month

Testing Procedure
Materials Provided
VITROS Chemistry Products CO2 DT Slides

Materials Required But Not Provided


VITROS Chemistry Products DT Calibrator Kit Quality control materials, such as VITROS Chemistry Products DT Control I & II VITROS Chemistry Products DT Reference Fluid VITROS DTE Dual Sample Cups VITROS DTE Pipette

Operating Instructions
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System. Bring all fluids and samples to room temperature, 18-28C (64-82F), prior to analysis. IMPORTANT:

Sample Dilution
Carbon dioxide concentrations outside the reportable (dynamic) range are not expected. Diluted samples should not be analyzed with VITROS CO2 DT Slides.

Calibration
Required Calibrators
VITROS Chemistry Products DT Calibrator Kit, bottles 1 and 2

Calibrator Preparation, Handling, and Storage


Refer to the instructions for use for VITROS DT Calibrator Kit.

Calibration Procedure
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Special Precautions
Calibrate CO2 in duplicate by running each calibrator bottle twice.

Version 1.0

Pub. No. C-308

VITQI

C0 2 DT
Carbon Dioxide When to Calibrate

INSTRUCTIONS FOR USE


Quality Control
Calibrate: When the slide lot number changes. When critical system parts are replaced due to service or maintenance. When government regulations require. - For example, in the USA, CLIA regulations require calibration or calibration verification at least once every six months. When the VITROS DT Reference Fluid lot number changes. The VITROS CO2 DT test may also need to be calibrated: If quality control results are consistently outside acceptable range. After certain service procedures have been performed. For additional information, refer to the operator's manual for your VITROS DT60/DT60II Chemistry System.

Calculations
The VITROS DT60/DT60 II Chemistry System measures the potential difference in millivolts between the two electrodes of a potentiometric slideone in contact with the sample to be analyzed and the other in contact with the electrolyte reference fluid. A linear relationship exists between the measured potential difference observed on the slide and the logarithm of carbon dioxide concentration, i.e. the Nernst equation for ion-selective electrodes. Once the calibration has been established for each slide lot, unknown carbon dioxide concentrations for a given sample can be determined using the software-resident math model and the measured potential difference.

Validity of a Calibration
Calibration parameters are automatically assessed by the VITROS DT60/DT60 II Chemistry System against a set of quality parameters resident within the analyzer software. Failure to meet any of the pre-defined quality parameters results in a failed calibration. The calibration report should be used in conjunction with quality control results to determine the validity of a calibration.

Reportable (Dynamic) Range Reportable (Dynamic) Range for CO; DT


Conventional and SI Units (mmol/L) 5-50

Traceability of the Calibration


Values assigned to the VITROS Chemistry Products DT Calibrator Kit for carbon dioxide are traceable to the Certified NIST (National Institute of Standards and Technology) Reference Material, SRM (Standard Reference Material) 192b. The Ortho-Clinical Diagnostics calibration laboratory uses SRM192b to calibrate the Corning Model 965 analyzer,9 lc utilizing thermal conductivity detection, to measure total carbon dioxide in support of carbon dioxide value assignment for the VITROS Chemistry Products DT Calibrator Kit.

Quality Control
Procedure Recommendations
I WARNING; Handle quality control materials as biohazardous material. Choose control levels that check the clinically relevant range. Analyze quality control materials in the same manner as patient samples, before or during patient sample processing. To verify system performance, analyze control materials: - After calibration. - According to local regulations or at least once each day that the test is being performed. - After specified service procedures are performed. Refer to the operator's manual for your VITROS DT60/DT60 II System. If control results fall outside your acceptable range, investigate the cause before deciding whether to report patient results. For general quality control recommendations, refer to Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition^ or other published guidelines. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Pub. No. C-308

Version 1.0

|*J VITRI

INSTRUCTIONS FOR USE


Expected Values and Reporting Units

COzDT
Carbon Dioxide

Quality Control Material Selection


IMPORTANT: VITROS DT Control I & II are recommended for use with the VITROS DT60/DT60 II Chemistry System. Evaluate the performance of other commercial control fluids for compatibility with this test before using for quality control.

Control materials other than VITROS DT Controls I & II may show a difference when compared with other Carbon Dioxide methods if they: - Depart from a true human matrix. - Contain high concentrations of preservatives, stabilizers, or other nonphysiological additives. Do not use control materials stabilized with ethylene glycol.

Quality Control Material Preparation and Storage


Refer to the instructions for use for VITROS Chemistry Products DT Control I & II or to other manufacturer's product literature.

Expected Values and Reporting Units


Reference Interval
This reference interval is the central 95% of results from a study of 60 apparently healthy adults from a working population (34 females and 26 males). Reference Interval for CO? DT Conventional and SI Units (mmol/L) 22-30 Each laboratory should confirm the validity of these intervals for the population it serves.

Reporting Units and Unit Conversion


The VITROS DT60/DT60 II Chemistry System may be programmed to report carbon dioxide results in conventional and SI units.

Reporting Units for CO, DT


Conventional and SI Units mmol/L

Limitations of the Procedure


Known Interferences
Lactate, hippurate, and other organic acids at significantly elevated concentrations have been reported to increase CO2 results.12 Diatrizoate sodium may increase CO2 results. Do not use specimens from patients receiving radiographic contrast agents containing this substance. The VITROS CO2 Slide method was screened for interfering substances following NCCLS Protocol EP7.13 The substances listed in the table, when tested at the concentrations indicated, caused the bias shown.

Known Interfering Substances for CO2 DT


Interferent* Bromide Iodide Nitrate Interferent Concentration Conv./SI Units (mmol/L)
2

1 3

Carbon Dioxide Cone. Conv./SI Units (mmol/L) 25 25 25

Average Bias Conv./SI Units (mmol/L) 2 4 11

It is possible that other interfering substances may be encountered. These results are representative; however, your results may differ somewhat due to test-to-test variation. The degree of interference at concentrations other than those listed might not be predictable.

Version 1.0

Pub. No. C-308

COzDT
Carbon Dioxide Other Limitations

INSTRUCTIONS FOR USE


Performance Characteristics
Certain drugs and clinical conditions are known to alter total carbon dioxide concentration in vivo. For additional information, refer to one of the published summaries.1415

Performance Characteristics
Method Comparison

The plot and table show the results of a comparison of samples analyzed on the VITROS DT60 II System with those analyzed using the VITROS 950 System. Testing followed NCCLS Protocol EP9.16

Method Comparison for CO2 DT: Serum


Conventional and SI Units
40 1 y =x

30 1
20

10

20

30

40

Comparative Method: VITROS 950 System (mmol/L)

Method Comparison for CO2 DT: Serum


Conventional and SI Units (mmol/L) n DT60 II System vs. 950 System 82 Slope 0.92 Correlation Coefficient 0.990 Range of Sample Cone. 13-38 Intercept Sy.x 2.13 0.82

Precision
Precision was evaluated with quality control materials on the VITROS DT60 II System following NCCLS Protocol EP5.17 The data presented are a representation of test performance and are provided as a guideline. Variables such as sample handling and storage, reagent handling and storage, laboratory environment, and system maintenance can affect reproducibility of test results.

Precision for CO2 DT: Serum


Conventional and SI Units (mmol/L) System VITROS DT60 II
* **

Mean Cone . 24 18

Within Day SD* Within Lab SD** Within Lab CV%** 0.6 0.5 0.7 0.6 3.1 3.2

No. Observ. 88 88

No. Days 22
22

Within Day precision was determined using two runs/day with two replications. Within Lab precision was determined using a single lot of slides and calibrating weekly.

Pub. No. C-308

Version 1.0

INSTRUCTIONS FOR USE


References

CO2DT
Carbon Dioxide

References
1. 2. NCCLS. Protection of Laboratory Workers from Instrument Biohazards and Infectious Diseases Transmitted by Blood, Body Fluids and Tissue; Approved Guideline. NCCLS Document M29 (OSBN 1-56238). NCCLS, Wayne, PA 19087; 1997. Doumas BT, et al. Differences Between Values for Plasma and Serum in Tests Performed in the Ektachem 700 XR Analyzer, and Evaluation of "Plasma Separator Tubes (PST)." Clin. Chem. 35:151-153; 1989. Calam RR. Specimen Processing Separator Gels: An Update. J Clin Immunoassay. 11:86-90; 1988. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Venipuncture. NCCLS Document H3. Wayne, PA:. NCCLS; 1991. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Skin Puncture. NCCLS Document H4. Wayne, PA: NCCLS;

1991.
HerrRD, Swanson T. Serum Bicarbonate Declines With Sample Size In Vacutainer Tubes. AJCP. 97:213-216; 1992. Slockbower JM, Blumenfeld TA (ed.). Collection and Handling of Laboratory Specimens. Philadelphia: Lippincott Co; 201; 1983. Clinical Laboratory Handbook for Patient Preparation and Specimen Handling. Fascicle VI: Chemistry/Clinical Microscopy. Northfield, IL: College of American Pathologists; 1992. Van Slyke D C , Neil! J.M. J. Biol. Chem., 1924; 61: 523-526. Corning 965 Carbon Dioxide Analyzer Instruction Manual, Corning Medical, Medfield, MA., 1977. NCCLS. Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition. NCCLS Document C24. Wayne, PA: NCCLS; 1999. Young DS. Effects of Preanalytical Variables on Clinical Laboratory Tests. Washington D.C.: AACC Press; 3-80; 1993. NCCLS. Interference Testing in Clinical Chemistry. NCCLS Document EP7. Wayne, PA: NCCLS; 1986. Young DS. Effects of Drugs on Clinical Laboratory Tests, ed. 4. Washington D.C.: AACC Press; 1995. Friedman RB, Young DS. Effects of Disease on Clinical Laboratory Tests. Washington, D.C.: AACC Press; 1990. NCCLS. Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline. NCCLS Document EP9. Wayne, PA:

9. 10, 11.
12. 13. 14. 15. 16. 17.

NCCLS; 1995.
NCCLS. User Evaluation of Precision Performance with Clinical Chemistry Devices. NCCLS Document EP5. Wayne, PA: NCCLS;

1992.

Glossary of Symbols
The Glossary of Symbols table defines the symbols used on the VITROS Chemistry Products packaging and on the VITROS Chemistry Products Assay Sheets.

Glossary of Symbols
Do Not Reuse Use by or Expiration Date (YYYY-MM-DD) Lot Number EC REP I Manufacturer Authorized Representative Contains Sufficient for "n" Tests For In Vitro Diagnostic Use Store At or Below Store At or Above Store Between Consult Instructions for Use Fragile, Handle with Care. Keep Dry This end up

SN
REF

Manufacturer's Serial Number Catalog Number or Product Code Attention: See Instructions for Use.

Version 1.0

Pub. No. C-308

C02DT
Carbon Dioxide

INSTRUCTIONS FOR USE


Revision History

Revision History
Date of Revision 2003-08-11 Description of Technical Changes* New format New organization and sections consistent with IVD Directive > Specimen Storage and Stability - updated stability > Reference Interval - updated data Limitations of the Procedure - added carbon dioxide concentration and bias for known interfering substances; removed the note Method Comparison - updated the data and plot > Precision - updated all data References - added all The change bars indicate the position of a technical amendment to the text with respect to the previous version of the document. Version 1.0

When this Instructions For Use is replaced, sign and date below and retain as specified by local regulations or laboratory policies, as appropriate.

Signature

Obsolete Date

C
Ortho-Clinical Diagnostics Mandeville House 62 The Broadway Amersham Buckinghamshire HP7 OHJ England Ortho-Clinical Diagnostics, Inc. 100 Indigo Creek Drive Rochester, NY 14626-5101

Ortho-Clinical Diagnostics
company

VITROS is a trademark of Ortho-Clinical Diagnostics, Inc. Ortho-Clinical Diagnostics, Inc., 2003. All rights reserved. Printed in USA.

Pub. No. C-308

Version 1.0

VITRCDS
Chemistry!

INSTRUCTIONS FOR USE


VITROS Chemistry Products CREA DT Slides Intended Use

CREAOT
Creatinine

For in vitro diagnostic use only. VITROS CREA DT Slides quantitatively measure creatinine (CREA) concentration in serum and plasma.

Summary and Explanation of the Test


Serum creatinine excretion is a function of lean body mass in normal persons and shows little or no response to dietary changes. The serum creatinine concentration is higher in men than in women. Serum creatinine is increased in acute or chronic renal failure, urinary tract obstruction, reduced renal blood flow, shock, dehydration, and rhabdomyolysis. Causes of low serum creatinine concentration include debilitation and decreased muscle mass. Exercise may cause an increased creatinine clearance.

Principles of the Procedure


The VITROS CREA DT Slide method is performed using the VITROS CREA DT Slide, the VITROS NH3 DT Slide, and the VITROS Chemistry Products DT Calibrator Kit on VITROS DT60/DT60 II Chemistry Systems. Refer to the VITROS NH3 DT Instructions for Use for an explanation of the principles of the procedure. The VITROS CREA DT Slide is a multilayered, analytical element coated on a polyester support. A drop of patient sample is deposited on the VITROS CREA DT Slide. The specimen is evenly distributed by the spreading layer to the underlying layers. Water and nonproteinaceous components travel to the underlying buffered reagent layer. Creatinine iminohydrolase catalyzes the hydrolysis of creatinine to produce ammonia. Both the ammonia produced in the previous reaction and endogenous ammonia from the sample diffuse through the semipermeable membrane to react with an ammonia indicator to produce a blue dye. After a fixed incubation time, the reflection density is proportional to the concentration of creatinine and endogenous ammonia in the sample. The software resident in the VITROS DT60/DT60 II System calculates a final creatinine concentration by subtracting the endogenous ammonia concentration measured by the VITROS NH3 DT Slide from the creatinine and endogenous ammonia concentration measured by the VITROS CREA DT Slide.

Reaction Sequence
creatinine + H2O
creatinine iminohydrolase

N-methylhydantoin + NH3 blue dye

NH3 + bromphenol blue (ammonia indicator)

Test Type and Conditions


Test Type and Conditions for CREA DT
Test Type Colorimetric VITROS DT60/DT60 II Module DT60/DT60 II Approximate Incubation Time 5 minutes Temperature 37C (98.6F). Wavelength 605 nm Sample Drop Volume 10 ML

Version 1.0

Pub. No. C-334

CREADT
Creatinine

INSTRUCTIONS FOR USE


Warnings and Precautions

Warnings and Precautions


For in vitro diagnostic use only. Take care when handling materials and samples of human origin. Since no test method can offer complete assurance that infectious agents are absent, consider all clinical specimens, controls, and calibrators potentially infectious. Handle specimens, solid and liquid waste, and test components in accordance with local regulations and NCCLS Guideline M29' or other published biohazard safety guidelines. For specific warnings and precautions for calibrators, quality control materials, and other components, refer to the instructions for use for the appropriate VITROS product, or to other manufacturer's product literature.

Reagents
Slide Diagram

Slide Ingredients
Reactive ingredients per cm
2
'*

Creatinine iminohydrolas'e (Bacillus species, EC.3.5.4.21) 0.5 U and bromphenol blue 27 ng.

y /,'' *
' ^' 4

Other ingredients
Pigment, binders, surfactants, buffer and stabilizer.

" ~"

1. Upper slide mount 2. Spreading layer (T1O2) 3. Reagent layer creatinine iminohydrojase buffer. pH 9.3 4. Semtpermeable membrane 6. indicator layer bromphenol blue 6. Support layer 7. Lower slide mount

, - - ' " "

Slide Handling
CAUTION: Do not use slides with damaged or incompletely sealed packaging.

Slide Preparation
IMPORTANT; The slide must reach room temperature, 18 "-28 C (64 ~82 F), before the wrapper is opened. Do not use unopened slides that have been at room temperature, 1 8 - 2 8 "C (64 "-82 F) for >48 hours. 1. 2. Remove the unopened slide from the box. Warm the unopened slide for 15 minutes. Unopened slides that remain in the sealed wrapper may be returned to refrigerator or freezer storage. 3. Remove the packaging and place the slide into the loading station within 15 minutes after opening.

Slide Storage and Stability


VITROS CREA DT Slides are stable until the expiration date on the carton when they are stored and handled as specified.

Slide Storage and Stability for CREA DT


Slides Unopened Storage Condition Room temperature 18-28 C (64-82F) Refrigerated 2-8C (36-46F) Frozen <-18C (<0F) Room temperature 18-28 'C (64-82 F) Stability <48 hours <4 weeks Until expiration date <15 minutes

Opened

Specimen Requirements
WARMING: Handle specimens as biohazardous material.

Specimens Recommended
Serum Plasma:2 EDTA Heparin (except ammonium heparin) Certain collection devices have been reported to affect other analytes and tests3 Confirm that your collection devices are compatible with this test.

IMPORTANT:

Pub. No. C-334

Version 1.0

INSTRUCTIONS FOR USI


Testing Procedure

CREADT
Creatinine

Serum and Plasma


Specimen Collection and Preparation
Collect specimens using standard laboratory procedures.4 5 Patient Preparation No special patient preparation is necessary.

Special Precautions NOTE: Avoid using ammonia-containing cleaning solutions or hand creams in the area around the analyzer.

Centrifuge specimens and remove the serum from the cellular material within 15 minutes of collection.2

Specimen Handling and Storage


Handle specimens as biohazardous material. Handle and store specimens in stoppered containers to avoid contamination and evaporation. Mix samples by gentle inversion and bring to room temperature, 18-28C (64-82F), prior to analysis. Two slides are used in the analysis of creatinine (the VITROS CREA DT Slide and the VITROS NH3 DT Slide). The VITROS NH3 DT Slide provides a blank correction value for normal concentrations of ammonia present in serum. Therefore, it is important to minimize the formation of ammonia during specimen storage.

Specimen Storage and Stability for CREA DT: Serum and Plasma2
Storage Room temperature Refrigerated Frozen Temperature 18-28C (64-82F) 2-8C (36-46F) <-18C (<0F) Stability not recommended <3 hours <24 hours

Testing Procedure
Materials Provided
. VITROS Chemistry Products CREA DT Slides

Materials Required But Not Provided


. . . . VITROS Chemistry Products NH3 DT Slides VITROS Chemistry Products DT Calibrator Kit Quality control materials, such as VITROS Chemistry Products DT Control I & II VITROS 7% BSA or reagent-grade water VITROS DT Pipette

Operating Instructions
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System. The VITROS NH3 DT Slide provides a blank correction value for normal concentrations of ammonia present in serum. The VITROS CREA DT Slide must be run first, followed by the VITROS NH3 DT Slide. The analyzer displays "INSERT NH3 SLIDE." When VITROS CREA DT or VITROS NH3 DT Slides are present in the incubator, the analyzer displays "ANALYZER READY-CREA/NH3 ONLY." No tests can be run until the CREA/NH3 results are complete. Bring all fluids and samples to room temperature, 18-28C (64-82F); analyze immediately. NOTE:

IMPORTANT:

Sample Dilution
If creatinine concentrations exceed the system's reportable (dynamic) range: 1. Dilute the sample with VITROS 7% BSA or reagent-grade water. 2. Reanalyze. 3. Multiply the result by the dilution factor to obtain an estimate of the original sample's creatinine concentration.

Version 1.0

Pub. No. C-334

CREA DT
Creatinine

INSTRUCTIONS FOR USE


Calibration

Calibration
Required Calibrators
VITROS Chemistry Products DT Calibrator Kit, bottles 1, 2, 3, and 4

Calibrator Preparation, Handling, and Storage


NOTE: To avoid ammonia formation, calibrator fluids should be prepared only when ready to calibrate and should be run within 1 hour after preparation. Because ammonia is produced in the VITROS BUN/UREA DT Slide reaction, the analyzer will display a warning message "ANALYER READYNO CREA/NH3" during calibration of VITROS BUN/UREA DT Slides. Therefore, it is recommended that creatinine and ammonia be the first tests calibrated after the preparation of calibrator fluids.

NOTE:

Refer to the instructions for use for VITROS DT Calibrator Kit.

Calibration Procedure
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

When to Calibrate
NOTE: The VITROS CREA DT test is dependent on correct calibration of the VITROS NH3 DT Slides used as blanks. Therefore, the VITROS NH3 DT Slides must be calibrated whenever VITROS CREA DT Slides are calibrated.

Calibrate: When the slide lot number changes. When critical system parts are replaced due to service or maintenance. When government regulations require. - For example, in the USA, CLIA regulations require calibration or calibration verification at least once every six months. The VITROS CREA DT test may also need to be calibrated: If quality control results are consistently outside acceptable range. After certain service procedures have been performed. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Calculations
Because the VITROS CREA DT Slide measures both creatinine and endogenous ammonia, the VITROS CREA DT Slide response is proportional to the concentration of both substances in the sample. A second slide, reactive only to ammonia, is sequentially measured for the blank correction of the VITROS CREA DT Slide response. Reflectance from both slides is measured at 605 nm after the fixed incubation time. Calibration using the blank-corrected calibration model consists of two partscalibration of the VITROS NH3 DT Slide (blank), followed by calibration of the VITROS CREA DT Slide. Once a calibration has been performed for each slide lot of VITROS NH3 DT Slides and VITROS CREA DT Slides, creatinine concentration in unknown samples can be determined using the software-resident endpoint colorimetric, blank-corrected math model and the responses obtained from both the VITROS CREA DT and VITROS NH3 DT Slides.

Validity of a Calibration
Calibration parameters are automatically assessed by the VITROS DT60/DT60 II Chemistry System against a set of quality parameters resident within the analyzer software. Failure to meet any of the pre-defined quality parameters results in a failed calibration. The calibration report should be used in conjunction with quality control results to determine the validity of a calibration.

Reportable (Dynamic) Range Reportable (Dynamic) Range for CREA DT


Conventional (mg/dL) SI Units <umol/L) 1-1326

0.01-15.0

For out-of-range samples, refer to "Sample Dilution."

Pub.

No. C-334

Version 1.0

INSTRUCTIONS FOR USE


Quality Control

CREA DT
Creatinine

Traceability of the Calibration


Values assigned to the VITROS Chemistry Products DT Calibrator Kit for creatinine are traceable to the Certified NIST (National Institute of Standards and Technology) Reference Material, SRM* (Standard Reference Material) 914a. The Ortho-Clinical Diagnostics calibration laboratory, uses SRM 914a to calibrate selected measurement procedures, including an HPLC (High Performance Liquid Chromatography)6 method and a rate Jaffe' method, to support creatinine value assignment for the VITROS DT Calibrator Kit.

Quality Control
Procedure Recommendations
| : Handle quality control materials as biohazardous material. Choose control levels that check the clinically relevant range. Analyze quality control materials in the same manner as patient samples, before or during patient sample processing. To verify system performance, analyze control materials: - After calibration. - According to local regulations or at least once each day that the test is being performed. - After specified service procedures are performed. Refer to the operator's manual for your VITROS DT60/DT60 II System. If control results fall outside your acceptable range, investigate the cause before deciding whether to report patient results. For general quality control recommendations, refer to Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition* or other published guidelines. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Quality Control Material Selection


MPORTANT: VITROS DT Control I & II are recommended for use with the VITROS DT60/DT60 II Chemistry System. Evaluate the performance of other commercial control fluids for compatibility with this test before using for quality control.

Control materials other than VITROS DT Controls I & II may show a difference when compared with other creatinine methods if they: - Depart from a true human matrix. - Contain high concentrations of preservatives, stabilizers, or other nonphysiological additives. Commercial control fluids that contain ammonia concentrations above the VITROS NH3 DT Slide reportable range (500 umol/L) will not allow a creatinine result to be calculated. Do not use control materials stabilized with ethylene glycol.

Quality Control Material Preparation and Storage


Refer to the instructions for use for VITROS Chemistry Products DT Control I & II or to other manufacturer's product literature.

Expected Values and Reporting Units


Reference Interval
These reference intervals are the central 95% of results from an internal study of apparently healthy adults from a working population (90 females, 105 males).

Reference Interval for CREA DT


Female Male Conventional Units (mg/dL) 0.7-1.2 mg/dL 0.8-1.5 mg/dL SI Units (umol/L) 62-106 umol/L 71-133 umol/L

Each laboratory should confirm the validity of these intervals for the population it serves.

Version 1.0

Pub. No. C-334

CREADT
Creatinine Reporting Units and Unit Conversion

INSTRUCTIONS FOR USE


Limitations of the Procedure

The VITROS DT60/DT60 II Chemistry System may be programmed to report creatinine results in conventional and SI units.

Reporting Units and Unit Conversion for CREA DT Conventional Units Sl Units
mg/dL Mmol/L (mg/dL x 88.4)

Limitations of the Procedure


Known Interferences
| Falsely elevated creatinine results have been reported for patients who have been given 5-fluorocytosine (flucytosine).9 An alternate method should be used to determine serum creatinine from these patients. Glucose at 600 mg/dL (33.3 mmol/L) may cause a decrease of 0.15 mg/dL (13.3 umol/L) in the creatinine result. VITROS CREA DT testing should not be done when any VITROS BUN/UREA DT Slides are in the incubator.

IMPORTANT: |

If a VITROS CREA DT Slide follows a VITROS BUN/UREA DT Slide immediately, high BUN values may increase creatinine values. A BUN value up to 40 mg/dL (14.3 mmol/L) may increase creatinine value by 0.3 mg/dL (26.6 umol/L) and an R18 code will be printed next to the creatinine result. Discard the result and repeat the sample without the VITROS BUN/UREA DT Slide in the incubator.

Other Limitations
Certain drugs and clinical conditions are known to alter creatinine concentration in vivo. For additional information, refer to one of the published summaries.10 "

Performance Characteristics
Method Comparison

The plots and table show the results of a comparison of samples analyzed on the VITROS DT60 II System with those analyzed using the HPLC comparative method.6

Method Comparison for CREA DT: Serum


Conventional Units IS 1400 1200 1000 800 600 400 200 0 12
Comparative Method: HPLC (mg/dL)

SI Units y =x

15

200 400 600 800 1000 1200 1400


Comparative Method: HPLC (Mmol/L)

Method Comparison for CREA DT: Serum


Conventional Units (mg/dL) n DT60 II System vs. comparative method 39 Correlation Slope Coefficient 1.03 0.992 Range of Sample Cone. Intercept Sy.x 0.04 0.44 SI Units (|jmol/L) Range of Sample Cone. 58-1017 Intercept 3.61 Sy.x 39.23

0.7-11.5

Pub. No. C-334

Version 1.0

INSTRUCTIONS FOR USE


References Precision

CREADT
Creatinine

Precision was evaluated with quality control materials on the VITROS DT60 II System following NCCLS Protocol EP5.12 The data presented are a representation of test performance and are provided as a guideline. Variables such as sample handling and storage, reagent handling and storage, laboratory environment, and system maintenance can affect reproducibility of test results.

Precision for CREA DT:Serum


Conventional Units (mg/dL) System VITROS DT60 II Mean Cone. 1.0 5.1 Within Day SD* 0.05 0.15 Within Lab S D " 0.06 0.20 Mean Cone. 93 455 SI Units (umol/L) Within Day SD* 4.1 13.5 Within Lab S D " 5.6 17.5 Within Lab CV%** 6.1 3.8 No. Observ. 83 84 No. Days 21 21

Within Day precision was determined using two runs/day with two replications. Within Lab precision was determined using a single lot of slides and calibrating weekly.

References
NCCLS. Protection of Laboratory Workers from Instrument Biohazards and Infectious Diseases Transmitted by Blood, Body Fluids and Tissue; Approved Guideline. NCCLS Document M29 (OSBN 1-56238). NCCLS, Wayne, PA 19087; 1997. 2. Clinical Laboratory Handbook for Patient Preparation and Specimen Handling. Fascicle VI: Chemistry/Clinical Microscopy. Northfield, IL: College of American Pathologists; 1992. 3. Calam RR. Specimen Processing Separator Gels: An Update. J Clin Immunoassay. 11:86-90; 1988. 4. NCCLS. Procedures forthe Collection of Diagnostic Blood Specimens by Venipuncture. NCCLS Document H3. Wayne, PA: NCCLS; 1991. 5. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Skin Puncture. NCCLS Document H4. Wayne, PA: NCCLS; 1991. 6. Ambrose RT, Ketchum DF, Smith JW. Creatinine Determined by "High Performance" Liquid Chromatography. Clin. Chem. 29: 256-259; 1983. 7. Jaffe M. Uber den Niederschlag welchen Pikrinsaure in normalen Harn erzeugt und uber eine neue Reaktion des Kreatinins. Z Physiol Chem. 10: 391-400; 1886. 8. NCCLS. Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition. NCCLS Document C24. Wayne, PA: NCCLS; 1999. 9. Mitchell RT, Marshall LH, Lefkowitz LB, Stratton CW. Falsely Elevated Serum Creatinine Levels Secondary to the Presence of 5Fluorocytosine. Am. J. Clin. Path. 84: 251-253; 1985. 10. Young DS. Effects of Drugs on Clinical Laboratory Tests, ed. 4. Washington D.C.: AACC Press; 1995. 11. Friedman RB, Young DS. Effects of Disease on Clinical Laboratory Tests. Washington, D.C.: AACC Press; 1990. 12. NCCLS. User Evaluation of Precision Performance with Clinical Chemistry Devices. NCCLS Document EP5. Wayne, PA: NCCLS; 1992. 1.

Glossary of Symbols
The Glossary of Symbols table defines the symbols used on the VITROS Chemistry Products packaging and on the VITROS Chemistry Products Assay Sheets.

Glossary of Symbols
Do Not Reuse Use by or Expiration Date (YYYY-MM-DD) Lot Number Manufacturer's Serial Number Catalog Number or Product Code Attention: See Instructions for Use. | REJ> J "\ v / V Manufacturer Authorized Representative Contains Sufficient for "n" Tests For In Vitro Diagnostic Use Store At or Below Store At or Above Store Between Consult Instructions for Use Fragile, Handle with Care. Keep Dry This end up

Version 1.0

Pub. No. C-334

CREADT
Creatinine

INSTRUCTIONS FOR USE


Revision History

Revision History
Date of Revision 2003-10-01 Version 1.0 Description of Technical Changes* > New format New organization and sections consistent with IVD Directive > Specimens Recommended - updated wording for heparin; added EDTA > Specimen Storage and Stability - updated stability values > Sample Dilution - added VITROS 7% BSA; removed isotonic saline > Reference Interval - updated data > Limitations of the Procedure - updated values > Method Comparison - updated all comparisons and the plots > Precision - updated all data > References - added all

The change bars indicate the position of a technical amendment to the text with respect to the previous version of the document.

When this Instructions For Use is replaced, sign and date below and retain as specified by local regulations or laboratory policies, as appropriate.

Signature

Obsolete Date

C
Ortho-Clinical Diagnostics Mandeville House 62 The Broadway Amersham Buckinghamshire HP7 OHJ England Ortho-Clinical Diagnostics, Inc. 100 Indigo Creek Drive Rochester, NY 14626-5101

' Ortho-Clinical Diagnostics


^eUmon company

VITROS is a trademark of Ortho-Clinical Diagnostics, Inc. Ortho-Clinical Diagnostics, Inc., 2003. All rights reserved. Printed in USA.

Pub. No. C-334

Version 1.0

(Products

VITRfflS
Chemistry!

INSTRUCTIONS FOR USE


VITROS Chemistry Products CRSC DT Slides Intended Use
For in vitro diagnostic use only. VITROS CRSC DT Slides quantitatively measure creatinine concentration in serum and plasma.

CRSCDT
Creatinine

Summary and Explanation of the Test


Serum creatinine excretion is a function of lean body mass in normal persons and shows little or no response to dietary changes. The serum creatinine concentration is higher in men than in women. Serum creatinine is increased in acute or chronic renal failure, urinary tract obstruction, reduced renal blood flow, shock, dehydration, and rhabdomyolysis. Causes of low serum creatinine concentration include debilitation and decreased muscle mass. Exercise may cause an increased creatinine clearance.

Principles of the Procedure


The VITROS CRSC DT Slide method is performed using the VITROS CRSC DT Slide and the VITROS Chemistry Products DT Calibrator Kit on VITROS DT60/DT60 II Chemistry Systems. The VITROS CRSC DT Slide is a multilayered, analytical element coated on a polyester support. A drop of patient sample is deposited on the slide and is evenly distributed by the spreading layer to the underlying layers. Creatinine diffuses to the reagent layer, where it is hydrolyzed to creatine in the rate-determining step. The creatine is converted to sarcosine and urea by creatine amidinohydrolase. The sarcosine, in the presence of sarcosine oxidase, is oxidized to glycine, formaldehyde, and hydrogen peroxide. The final reaction involves the peroxidase-catalyzed oxidation of a leuco dye to produce a colored product. Following addition of the sample, the slide is incubated. During the initial reaction phase, endogenous creatine in the sample is oxidized. The rate of change in reflection density is proportional to the concentration of creatinine present in the sample.

Reaction Sequence
creatinine + H2O creatine + H2O sarcosine + O2 + H2O H2O2 + leuco dye
creatinine amidohydrolase

creatine sarcosine + urea glycine + formaldehyde + H2O2 dye + 2H2O

creatine amidinohydrolase

sarcosine oxidase peroxidase

Test Type and Conditions


Test Type and Conditions for CRSC DT
Test Type Rate VITROS DT60/DT60 II Module DTSC/DTSC II Approximate

Incubation Time
5 minutes

Temperature 37C (98.6F)

Wavelength 680 nm

Sample Drop Volume 10 ML

Version 1.0

Pub. No. C-353

CRSC or
Creatinine

INSTRUCTIONS FOR USE


Warnings and Precautions

Warnings and Precautions


For in vitro diagnostic use only. Take care when handling materials and samples of human origin. Since no test method can offer complete assurance that infectious agents are absent, consider all clinical specimens, controls, and calibrators potentially infectious. Handle specimens, solid and liquid waste, and test components in accordance with local regulations and NCCLS Guideline M291 or other published biohazard safety guidelines. For specific warnings and precautions for calibrators, quality control materials, and other components, refer to the instructions for use for the appropriate VITROS product, or to other manufacturer's product literature.

Reagents
Slide Diagram

Slide Ingredients
Reactive ingredients per cm
2

Creatinine amidohydrolase (Flavobacterium sp., E.C.3.5.2.10) 0.20 U; creatine amidinohydrolase {Flavobacterium sp., E.C.3.5.3.3) 4.7 U; sarcosine oxidase (Bacillus sp., E.C.1.5.3.1) 0.55 U; peroxidase (horseradish root, E.C.1.11.1.7) 1.6 U; and 2-(3,5-dimethoxy-4-hydroxyphenyl)-4,5-bis(4-dimethylaminophenyl) imidazole (leuco dye) 32 ng.

1. Upper slide mount 2. Spreading layer (TIO2) 3. Reagent layer creatinine amidohydrolase creatine amidinohydrolase sarcosine oxidase peroxidase leuco dye buffer, pH 7.0

4. Support layer
5. Lower slide mount

Other ingredients
Pigment, binders, surfactants, stabilizer, scavenger, chelator, buffer, dye solubilizer and cross-linking agent.

Slide Handling
Do not use slides with damaged or incompletely sealed packaging.

Slide Preparation
IMPORTANT; The slide must reach room temperature, 18-28C (64-82F), before the wrapper is opened. Do not use unopened slides that have been at room temperature, 18 "-28 C (64-82F) for >48 hours. 1. 2. Remove the unopened slide from the box. Warm the unopened slide for 15 minutes. Unopened slides that remain in the sealed wrapper may be returned to refrigerator or freezer storage. 3. Remove the packaging and place the slide into the loading station within 15 minutes after opening.

Slide Storage and Stability


VITROS CRSC DT Slides are stable until the expiration date on the carton when they are stored and handled as specified.

Slide Storage and Stability for CRSC DT


Slides Unopened Opened Storage Condition Room temperature 18-28C (64-82F) Frozen <-18C (<0F) Room temperature 18-28C (64-82F) Stability <48 hours Until expiration date <15 minutes

Pub. No. C-353

Version 1.0

INSTRUCTIONS FOR USE


Specimen Requirements

CRSGDT
Creatinine

Specimen Requirements
Handle specimens as biohazardous material.

Specimens Recommended

Serum Plasma:2 MPORTANT:

Heparin Certain collection devices have been reported to affect other analytes and tests3 Confirm that your collection devices are compatible with this test.

Specimens Not Recommended


Do not use specimens obtained through catheters used to infuse hyperalimentation fluid. Refer to "Limitations of the Procedure."

Serum and Plasma Specimen Collection and Preparation


Collect specimens using standard laboratory procedures.4 5 Patient Preparation No special patient preparation is necessary. Special Precautions Centrifuge specimens and remove the serum or plasma from the cellular material within 4 hours of collection.6 Specimen Handling and Storage Handle and store specimens in stoppered containers to avoid contamination and evaporation. Mix samples by gentle inversion and bring to room temperature, 18-28C (64-82F), prior to analysis.

Specimen Storage and Stability for CRSC DT: Serum and Plasma6 Temperature Storage Stability
Room temperature Refrigerated Frozen 18-28OC(64-82OF) 2-8C (36-46F) <-18C(<0F) <5 days <30 days <lndefinite

Testing Procedure
Materials Provided
VITROS Chemistry Products CRSC DT Slides

Materials Required But Not Provided


. . VITROS Chemistry Products DT Calibrator Kit Quality control materials, such as VITROS Chemistry Products DT Control I & II VITROS Chemistry Products 7% BSA Isotonic saline VITROS DT Pipette

Operating Instructions
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System. Bring all fluids and samples to room temperature, 18-28C (64-82F); analyze immediately. IMPORTANT:

Sample Dilution
If creatinine concentrations exceed the system's reportable (dynamic) range or if the analyzer displays an L-11 or L-13 error code (indicating high background density, usually due to an elevated creatine concentration): 1. Dilute the sample with 7% BSA or isotonic saline. 2. Reanalyze. 3. Multiply the results by the dilution factor to obtain an estimate of the original sample's creatinine concentration.

Version 1.0

Pub. No. C-353

CRSCDT
Creatinine

INSTRUCTIONS FOR USE


Calibration

Calibration
Required Calibrators
VITROS Chemistry Products DT Calibrator Kit, bottles 1, 2, and 4

Calibrator Preparation, Handling, and Storage


Refer to the instructions for use for VITROS DT Calibrator Kit.

Calibration Procedure
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

When to Calibrate
Calibrate: When the slide lot number changes. When critical system parts are replaced due to service or maintenance. When government regulations require. - For example, in the USA, CLIA regulations require calibration or calibration verification at least once every six months. The VITROS CRSC DT test may also need to be calibrated: If quality control results are consistently outside acceptable range. After certain service procedures have been performed. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Calculations
Based on sequential readings of the slide's reflectance at 680 nm over the defined incubation period, a rate of change in reflectance is determined. This rate is used in the software-resident multi-point rate calibration model to compute enzyme activity. Once a calibration has been performed for each slide lot, creatinine concentration in unknown samples can be determined from the rate of change in reflectance measured for each unknown test slide.

Validity of a Calibration
Calibration parameters are automatically assessed by the VITROS DT60/DT60 II Chemistry System against a set of quality parameters resident within the analyzer software. Failure to meet any of the pre-defined quality parameters results in a failed calibration. The calibration report should be used in conjunction with quality control results to determine the validity of a calibration.

Reportable (Dynamic) Range Reportable (Dynamic) Range for CRSC DT


Conventional (mg/dL) SI Units (umol/L) 4-1459

0.1-16.5

For out-of-range samples, refer to "Sample Dilution."

Traceability of the Calibration


Values assigned to the VITROS Chemistry Products DT Calibrator Kit for creatinine are traceable to the Certified NIST (National Institute of Standards and Technology) Reference Material, SRM (Standard Reference Material) 914a. The Ortho-Clinical Diagnostics calibration laboratory, uses SRM 914a to calibrate selected measurement procedures, including an HPLC (High Performance Liquid Chromatography)7 method and a rate Jaffe8 method, to support creatinine value assignment for the VITROS DT Calibrator Kit.

Quality Control
Procedure Recommendations
| " Handle quality control materials as biohazardous material. Choose control levels that check the clinically relevant range. Analyze quality control materials in the same manner as patient samples, before or during patient sample processing. To verify system performance, analyze control materials: - After calibration. - According to local regulations or at least once each day that the test is being performed. - After specified service procedures are performed. Refer to the operator's manual for your VITROS DT60/DT60 II System.

Pub. No. C-353

Version 1.0

INSTRUCTIONS FOR USE


Expected Values and Reporting Units

CRSC DT
Creatinine

If control results fall outside your acceptable range, investigate the cause before deciding whether to report patient results. For general quality control recommendations, refer to Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition9 or other published guidelines. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Quality Control Material Selection


IMPORTANT: VITROS DT Control I & II are recommended for use with the VITROS DT60/DT60 II Chemistry System. Evaluate the performance of other commercial control fluids for compatibility with this test before using for quality control.

Control materials other than VITROS DT Controls I & II may show a difference when compared with other creatinine methods if they: - Depart from a true human matrix. - Contain high concentrations of preservatives, stabilizers, or other nonphysiological additives. Liquid serum often contain high creatine levels and may give L-11 or L-13 error codes. Do not use control materials stabilized with ethylene glycol.

Quality Control Material Preparation and Storage


Refer to the instructions for use for VITROS Chemistry Products DT Control I & I or to other manufacturer's product literature.

Expected Values and Reporting Units


Reference Interval
These reference intervals are the central 95% of results from an internal study of apparently healthy adults from a working population (105 males, 90 females).

Reference Interval for CRSC DT


Conventional Units (mg/dL) Male Female

0.8-1.5 0.7-1.2

SI Units (umol/L) 71-133 62-106

Each laboratory should confirm the validity of these intervals for the population it serves.

Reporting Units and Unit Conversion


The VITROS DT60/DT60 II Chemistry System may be programmed to report creatinine results in conventional and SI units.

Reporting Units and Unit Conversion for CRSC DT Conventional Units SI Units
mg/dL umol/L (mg/dL x 88.4)

Limitations of the Procedure


Known Interferences
Creatine: At a creatinine concentration of 1.5 mg/dL (133 umol/L), creatine greater than 8 mg/dL (707 umol/L) will be flagged with an L-11 error code (because highly elevated creatine concentrations may cause excessive background density). For unflagged samples, residual bias because of creatine will be less than 0.15 mg/dL (13 umol/L) at low creatinine concentrations. Residual bias for unflagged samples will be less than 4% at high creatinine concentrations. Refer to "Sample Dilution" for dilution instructions. Proline: Patients receiving hyperalimentation fluids containing proline may show an increase of 0.2 mg/dL (18 umol/L). Do not collect specimens from intravenous fluid lines contaminated with hyperalimentation fluid. Dobutamine: Specimens contaminated with dobutamine from intravenous fluid have been reported to show a significant negative bias. A dobutamine concentration of 83 ug/mL caused a decrease of 2.7 mg/dL (239 umol/L) from an initial creatinine concentration of 4.8 mg/dL (424 umol/L).10 Lidocaine: Patients on long-term lidocaine therapy may show an increase of up to 1.0 mg/dL (88 umol/L) due to a metabolite of lidocaine, N-ethyl glycine (NEG).11

Version 1.0

Pub. No. C-353

CRSC DT
Creatinine

INSTRUCTIONS FOR USE


Performance Characteristics

The VITROS CRSC DT Slide method was screened for interfering substances. The substances listed in the table, when tested at the concentrations indicated, caused the bias shown.

Known Interfering Substances for CRSC DT


Interferent* Dipyrone (Metamizol) Interferent Concentration Creatinine Concentration SI (umol/L) Conv. (mg/dL) Average Bias SI (|jmol/L) Conv. (mg/dL) -0.6 (-53) (-35 umol/L) -0.4 mg/dL

40mg/dL (1138 Mmol/L) 1.0 (88) (115 pmol/L) /V-acetylcysteine 90 mg/dL (5.50 mmol/L) 1.3 mg/dL * It is possible that other interfering substances may be encountered. These results are representative; however, your results may differ somewhat due to test-to-test variation. The degree of interference at concentrations other than those listed might not be predictable.

Other Limitations
Certain drugs and clinical conditions are known to alter creatinine concentration in vivo. For additional information, refer to one of the published summaries.12 "

Performance Characteristics
Method Comparison

The plots and table show the results of a comparison of the VITROS DT60 II Chemistry System with the VITROS 950 Chemistry System.

Method Comparison for CRSC DT: Serum Conventional Units


IX 15 12

SI Units 1500 1200 900 y =x

y =x

9
D w O

1
O

600 300 0

12

15

IS

300

600

900

1200

1500

Comparative Method: VITROS 950 System (mg/dL)

Comparative Method: VITROS 950 System (Mmol/L)

Method Comparison for CRSC DT: Serum


Conventional Units (mg/dL) n DT60 II System vs. comparative method 70 Correlation Slope Coefficient 0.97 0.999 Range of Sample Cone. 0.7-12.2 Intercept Sy.x 0.1 0.19 SI Units (Mmol/L) Range of Sample Cone. 58-1077 Intercept 8.86 Sy.x 16.72

Pub. No. C-353

Version 1.0

INSTRUCTIONS FOR USE


References

CRSCDT
Creatinine

Precision
Precision was evaluated with quality control materials on VITROS the DTSC II System following NCCLS Protocol EP5.'4 The data presented are a representation of test performance and are provided as a guideline. Variables such as sample handling and storage, reagent handling and storage, laboratory environment, and system maintenance can affect reproducibility of test results.

Precision for CRSC DT:Serum


Conventional Units (mg/dL) System VITROS DT60 II Mean Cone. 1.2 5.9 Within Day SD* 0.01 0.06 Within Lab SD** 0.02 0.10 SI Units (umol/L) Mean Cone. 110 521 Within Day SD* 1.1 5.1 Within Lab SD** 1.5 8.8 Within Lab CV%** 1.3 1.7 No. Observ. 88 88 No. Days 22 22

Within Day precision was determined using two runs/day with two replications. Within Lab precision was determined using a single lot of slides and calibrating weekly.

References
1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. NCCLS. Protection of Laboratory Workers from Instrument Biohazards and Infectious Diseases Transmitted by Blood, Body Fluids and Tissue; Approved Guideline. NCCLS Document M29 (OSBN 1-56238). NCCLS, Wayne, PA 19087; 1997. Doumas BT, et al. Differences Between Values for Plasma and Serum in Tests Performed in the Ektachem 700 XR Analyzer, and Evaluation of "Plasma Separator Tubes (PST)." Clin. Chem. 35:151-153; 1989. Calam RR. Specimen Processing Separator Gels: An Update. J Clin Immunoassay. 11:86-90; 1988. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Venipuncture. NCCLS Document H3. Wayne, PA: NCCLS; 1991. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Skin Puncture. NCCLS Document H4. Wayne, PA: NCCLS; 1991. Clinical Laboratory Handbook for Patient Preparation and Specimen Handling. Fascicle VI: Chemistry/Clinical Microscopy. Northfield, IL: College of American Pathologists; 1992. Ambrose RT, Ketchum DF, Smith JW. Creatinine Determined by "High Performance" Liquid Chromatography. Clin. Chem. 29: 256-259; 1983. Jaffe M. Uber den Niederschlag welchen Pikrinsaure in normalen Harn erzeugt und uber eine neue Reaktion des Kreatinins. Z Physiol Chem. 10:391-400; 1886. NCCLS. Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition. NCCLS Document C24. Wayne, PA: NCCLS; 1999. DalyT, Kempe K, Scott M. "Bouncing" Creatinine Levels. NEJM. 334(26): 1749; 1996. Sena SF, Syed D, Romeo R, Krzymowski GA, McComb RB. Lidocaine Metabolite and Creatinine Measurements in the Ektachem 700: Steps to Minimize its Impact on Patient Care. Clin. Chem. 34:10; 1988. Young DS. Effects of Drugs on Clinical Laboratory Tests, ed. 4. Washington D.C.: AACC Press; 1995. Friedman RB, Young DS. Effects of Disease on Clinical Laboratory Tests. Washington, D.C.: AACC Press; 1990. NCCLS. User Evaluation of Precision Performance with Clinical Chemistry Devices. NCCLS Document EP5. Wayne, PA: NCCLS; 1992.

Glossary of Symbols
The Glossary of Symbols table defines the symbols used on the VITROS Chemistry Products packaging and on the VITROS Chemistry Products Assay Sheets.

Glossary of Symbols
Do Not Reuse Use by or Expiration Date (YYYY-MM-DD) Lot Number Manufacturer's Serial Number Catalog Number or Product Code Attention: See Instructions for Use. [ EC [ BEP | \ / \ / Manufacturer Authorized Representative Contains Sufficient for "n" Tests For In Vitro Diagnostic Use Store At or Below Store At or Above Store Between Consult Instructions for Use Fragile, Handle with Care. Keep Dry This end up

Version 1.0

Pub. No. C-353

VITR^IS 0

CRSC DT
Creatinine

INSTRUCTIONS FOR USE


Revision History

Revision History
Date of Revision 2003-04-30 Version 1.0 Description of Technical Changes* New format New organization and sections consistent with IVD Directive Specimen Storage and Stability - updated values for room temperature and refrigerated Materials Required But Not Provided - added VITROS 7% BSA; deleted isotonic saline Reportable (Dynamic) Range - corrected SI units Quality Control Material Selection - removed the statement regarding Tris buffer Limitations of the Procedure - updated information and corrected creatinine concentration in SI units Method Comparison - updated all data and plots Precision - updated all data References - added all

The change bars indicate the position of a technical amendment to the text with respect to the previous version of the document.

When this Instructions For Use is replaced, sign and date beiowand retain as specified by local regulations or laboratory policies, as appropriate.

Signature

Obsolete Date

C
Ortho-Clinical Diagnostics Mandeville House 62 The Broadway Amersham Buckinghamshire HP7 OHJ England Ortho-Clinical Diagnostics, Inc. 100 Indigo Creek Drive Rochester, NY 14626-5101

' Ortho-Clinical Diagnostics


weH company

VITROS is a trademark of Ortho-Clinical Diagnostics, Inc. Ortho-Clinical Diagnostics, Inc., 2003. All rights reserved. Printed in USA.

Pub. No. C-353

Version 1.0

Prodwcts

Chemistry

INSTRUCTIONS FOR OSE


VITROS Chemistry Products Fe DT Slides Intended Use
For in vitro diagnostic use only. VITROS Fe DT Slides quantitatively measure iron (Fe) concentration in serum and plasma.

FeDT
Iron

Summary and Explanation of the Test


Most body iron is found in hemoglobin. The serum measurement of iron is useful in the differential diagnosis of anemia, iron deficiency anemia, thalassemia, possible sideroblastic anemia, and iron poisoning. Serum iron is increased in hemosiderosis, hemolytic anemias, thalassemia, sideroblastic anemias, hepatitis, acute hepatic necrosis, hemochromatosis, inappropriate iron therapy, and iron poisoning. Serum iron is decreased in cases of insufficient dietary iron, chronic blood loss, inadequate absorption of iron, impaired release of iron stores (commonly observed in inflammation), infection, and chronic diseases.1

Principles of the Procedure


The VITROS Fe DT Slide method is performed using the VITROS Fe DT Slide and the VITROS Chemistry Products DT Calibrator Kit on VITROS DT60 II Chemistry Systems. The VITROS Fe DT Slide is a multilayered, analytical element coated on a polyester support. A drop of patient sample is deposited on the slide and is evenly distributed by the spreading layer to the underlying layers. Iron (as ferric ion) is removed from transferrin at acidic pH and migrates to the reducing layer, where ascorbic acid reduces iron to the ferrous form. The ferrous ion then is bound to the dye and forms a colored complex in the reagent layer. The rate of change in reflection density is proportional to the iron concentration in the sample.

Reaction Sequence
transferrin - Fe* Fe + ascorbic acid
pH4.0

- > transferrin + Fe+: ->Fe+ - > Fe+2 - dye (colored complex)

Fe+2 + dye

Test Type and Conditions


Test Type and Conditions for Fe DT
Test Type Rate VITROS DT60 II Module DTSC II Approximate Incubation Time 5 minutes Temperature 37C (98.6F) Wavelength 630 nm Sample Drop Volume 10 uL

Warnings and Precautions


For in vitro diagnostic use only. Take care when handling materials and samples of human origin. Since no test method can offer complete assurance that infectious agents are absent, consider all clinical specimens, controls, and calibrators potentially infectious. Handle specimens, solid and liquid waste, and test components in accordance with local regulations and NCCLS Guideline M292 or other published biohazard safety guidelines. For specific warnings and precautions for calibrators, quality control materials, and other components, refer to the instructions for use for the appropriate VITROS product, or to other manufacturer's product literature.

Version 1.0

Pub. No. C-371

FeDT
Iron

INSTRUCTIONS FOR USE


Reagents

Reagents
Slide Diagram

Slide Ingredients
Reactive ingredients per cm
2

-/'

y'

Ascorbic acid 160 ug; and N-(4-(2,4-bis(1,1-dimethylpropyl) phenoxy)butyl)5-methoxy-6((2,3,6,7-tetrahydro-8-1H,5H-benzo-(ij)-quinolizin-9-yl)azo)3-pyridine sulfonamide (dye) 5 ug.

'3

Other ingredients
Binders, buffer, pigment, surfactants, stabilizer, chelator, dye solubilizer and cross-linking agent.

- s
$

1. Upper slide mount 2. Spreading layer (BaSOit) 3. Reducing layer ascorbic acid 4. Reagent layer buffer, pH 4.0 dye 6. Support layer 6. Lower slide mount

Slide Handling
Do not use slides with damaged or incompletely sealed packaging.

Slide Preparation
IMPORTANT: The slide must reach mom temperature, 18-28C (64-82F), before the wrapper is opened. Do not use unopened slides that have been at room temperature, 18-28C (64 -82 F) for >48 hours. 1. 2. Remove the unopened slide from the box. Warm the unopened slide for 15 minutes. Unopened slides that remain in the sealed wrapper may be returned to refrigerator or freezer storage. 3. Remove the packaging and place the slide into the loading station within 15 minutes after opening.

Slide Storage and Stability


VITROS Fe DT Slides are stable until the expiration date on the carton when they are stored and handled as specified.

Slide Storage and Stability for Fe DT Slides Storage Condition


Unopened Room temperature Refrigerated Frozen Room temperature 18-28C (64-82F) 2-8C (36-46F) <-18C(<0F) 18-28C (64-82F)

Opened

Stability <48 hours Until expiration date Until expiration date <15 minutes

Specimen Requirements
Handle specimens as biohazardous material.

Specimens Recommended
Serum Plasma: Heparin Certain collection devices have been reported to affect other analytes and tests.3 Confirm that your collection devices are compatible with this test.

IMPORTANT;

Specimens Not Recommended


Do not use hemolyzed specimens because of the high concentration of iron in hemoglobin.1

Pub.

No. C-371

Version 1.0

[] VITRI

INSTRUCTIONS FOR USE


Testing Procedure Serum and Plasma
Specimen Collection and Preparation
Collect specimens using standard laboratory procedures.4 5 Patient Preparation No special patient preparation is necessary. Special Precautions Centrifuge specimens and remove the serum or plasma from the cellular material within 4 hours of collection.6

FeDT
Iron

Specimen Handling and Storage


Handle specimens as biohazardous material. Handle and store specimens in stoppered containers to avoid contamination and evaporation. Mix samples by gentle inversion and bring to room temperature, 18-28C (64-82F), prior to analysis.

Specimen Storage and Stability for Fe DT: Serum and Plasma6 Temperature Stability Storage Room temperature <4 days 18-28C (64-82F) Refrigerated <7 days 2-8C (36-46F) Frozen <-18C(<0F) <3 months

Testing Procedure
Materials Provided
VITROS Chemistry Products Fe DT Slides

Materials Required But Not Provided


VITROS Chemistry Products DT Calibrator Kit Quality control materials, such as VITROS Chemistry Products DT Control I & II Iron-free reagent-grade water VITROS DT Pipette

Operating Instructions
Refer to the operator's manual for your VITROS DT60 II Chemistry System. Bring all fluids and samples to room temperature, 18-28C (64-82F), prior to analysis.

I
Sample Dilution

If iron concentrations exceed the system's reportable (dynamic) range: 1. Dilute the sample with iron-free reagent-grade water. 2. Reanalyze. 3. Multiply the results by the dilution factor to obtain an estimate of the original sample's Iron concentration.

Calibration
Required Calibrators
VITROS Chemistry Products DT Calibrator Kit, bottles 1, 2, and 4

Calibrator Preparation, Handling, and Storage


Refer to the instructions for use for VITROS DT Calibrator Kit.

Calibration Procedure
Refer to the operator's manual for your VITROS DT60 II Chemistry System.

Version 1.0

Pub. No. C-371

FeDT
Iron

INSTRUCTIONS FOR USE


Quality Control
Calibrate: When the slide lot number changes. When critical system parts are replaced due to service or maintenance. When government regulations require. - For example, in the USA, CLIA regulations require calibration or calibration verification at least once every six months. The VITROS Fe DT test may also need to be calibrated: If quality control results are consistently outside acceptable range. After certain service procedures have been performed. For additional information, refer to the operator's manual for your VITROS DT60 II Chemistry System.

When to Calibrate

Calculations
Reflectance from the slide is read at 630 nm during the incubation period, and the rate of change in reflectance is calculated. Once a calibration has been performed for each slide lot, iron concentration in unknown samples can be determined using the software-resident two-point rate math model and the change in reflectance calculated for each unknown test slide.

Validity of a Calibration
Calibration parameters are automatically assessed by the VITROS DT60 II Chemistry System against a set of quality parameters resident within the analyzer software. Failure to meet any of the pre-defined quality parameters results in a failed calibration. The calibration report should be used in conjunction with quality control results to determine the validity of a calibration.

Reportable (Dynamic) Range Reportable (Dynamic) Range for Fe DT


Conventional (ug/dL) SI Units ((jmol/L) 1.8-89.6

10-500

For out-of-range samples, refer to "Sample Dilution."

Traceability of the Calibration


Values assigned to the VITROS Chemistry Products DT Calibrator Kit for iron are traceable to the Certified NIST (National Institute of Standards and Technology) Reference Material, SRM" (Standard Reference Material) 937. The Ortho-Clinical Diagnostics (OCD) calibration laboratory uses SRM" 937 to calibrate the proposed NCCLS standard iron method,7 modified according to the International Committee for Standardization in Hematology (ICSH) recommendation to use ferene dye,8 to support iron value assignment for the VITROS DT Calibrator Kit.

Quality Control
Procedure Recommendations
| Handle quality control materials as biohazardous material. Choose control levels that check the clinically relevant range. Analyze quality control materials in the same manner as patient samples, before or during patient sample processing. To verify system performance, analyze control materials: - After calibration. - According to local regulations or at least once each day that the test is being performed. - After specified service procedures are performed. Refer to the operator's manual for your VITROS DT60 II System. If control results fall outside your acceptable range, investigate the cause before deciding whether to report patient results. For general quality control recommendations, refer fo Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition9 or other published guidelines. For additional information, refer to the operator's manual for your VITROS DT60 II Chemistry System.

Pub. No. C-371

Version 1.0

INSTRUCTIONS FOR USE


Limitations of the Procedure

FeDT
Iron

Quality Control Material Selection


IMPORTANT: VITROS DT Control I & II are recommended for use with the VITROS DT60 II Chemistry System. Evaluate the performance of other commercial control fluids for compatibility with this test before using for quality control.

Control materials other than VITROS DT Controls I & II may show a difference when compared with other Iron methods if they: - Depart from a true human matrix. - Contain high concentrations of preservatives, stabilisers, or other nonphysiological additives. Do not use control materials stabilized with ethylene glycol.

Quality Control Material Preparation and Storage


Refer to the instructions for use for VITROS Chemistry Products DT Control I & II or to other manufacturer's product literature.

Reference Interval
These reference intervals are the central 95% of results from an internal study of 529 apparently healthy adults from a working population (382 females and 147 males).

Reference Interval for Fe DT


Conventional Units (Mg/dL) Females Males SI Units (umol/L) 6.6-30.4 8.8-32.4

37-170 49-181

Each laboratory should confirm the validity of these intervals for the population it serves.

Reporting Units [and Unit Conversion]


The VITROS DT60 II Chemistry System may be programmed to report Fe results in conventional and SI units.

Reporting Units and Unit Conversion for Fe DT


Conventional Units
Mg/dL

SI Units Mmol/L (ug/dL x 0.1791)

Limitations of the Procedure


Known Interferences
The VITROS Fe DT Slide method was screened for interfering substances following NCCLS Protocol EP7.10 The substances listed in the table, when tested at the concentrations indicated, caused the bias shown. Desferol (Deferoxamine Mesylate) at a concentration of 250 mg/dL and higher results in iron concentrations below the system range. Imferon produces a positive iron bias of 5 ,ug/dL for each 100 i-ig/dL of imferon.

Other Limitations
Certain drugs and clinical conditions are known to alter Iron concentration in vivo. For additional information, refer to one of the published summaries.11'12

Version 1.0

Pub. No. C-371

FeDT
Iron

INSTRUCTIONS FOR USE


Performance Characteristics

Performance Characteristics
Method Comparison

The plots and table show the results of a comparison of samples analyzed on the VITROS DT60 II System with those analyzed using the VITROS 950 System. Testing followed NCCLS Protocol EP9.13

Method Comparison for Fe DT: Serum Conventional Units


600 500 "
E

SI Units

y=x

100

80 60

400 300 '


2O0

8
100 "

20

0
100 200

300

400

500

600

20

40

60

80

too

Comparative Method: VITROS 950 (Mg/dL)

Comparative Method: VITROS 950 (Mmol/L)

Method Comparison for Fe DT: Serum


Conventional Units (pg/dL) n DT60 II System vs. comparative method 53 Correlation Slope Coefficient 1.03 0.999 Range of Sample Cone. 19-448 Intercept Sy.x -1.47 4.71 SI Units (umol/L) Range of Sample Cone. 3.5-80.2 Intercept Sy.x -0.26 0.84

Precision
Precision was evaluated with quality control materials on the VITROS DT60 II System following NCCLS Protocol EP5.14 The data presented are a representation of test performance and are provided as a guideline. Variables such as sample handling and storage, reagent handling and storage, laboratory environment, and system maintenance can affect reproducibility of test results.

Precision for Fe DT: Serum


Mean Cone. 72 240 Conventional Units (Mg/dL) Within Within Day SD* Lab SD** 1.6 4.6 2.2 6.7 Mean Cone. 12.8 43.0 SI Units (Mmol/L) Within Within Day SD* Lab SD** 0.28 0.83 0.39 1.20 Within Lab CV%** 3.0 2.8 No. Observ. 88 88 No. Days 22 22

System VITROS DT60 II

Within Day precision was determined using two runs/day with two replications. Within Lab precision was determined using a single lot of slides and calibrating weekly.

Pub. No. C-371

Version 1.0

INSTRUCTIONS FOR USE


References

FeDT
Iron

References
1. .2. 3. 4. 5. 6. 7. 8. 9. Tietz NW(ed). Fundamentals of Clinical Chemistry, ed. 3. Philadelphia: WB Saunders; 819-821; 1987. NCCLS. Protection of Laboratory Workers from Instrument Biohazards and Infectious Diseases Transmitted by Blood, Body Fluids and Tissue; Approved Guideline. NCCLS Document M29 (OSBN 1-56238). NCCLS, Wayne, PA 19087; 1997. Calam RR. Specimen Processing Separator Gels: An Update. J Clin Immunoassay. 11:86-90; 1988. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Venipuncture. NCCLS Document H3. Wayne, PA: NCCLS;

1991.
NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Skin Puncture. NCCLS Document H4. Wayne, PA: NCCLS; 1991. Clinical Laboratory Handbook for Patient Preparation and Specimen Handling. Fascicle VI: Chemistry/Clinical Microscopy. Northfield, IL: College of American Pathologists; 1992. NCCLS. The Determination of Serum Iron and Total iron binding-capacity; Proposed Standard. NCCLS Document H17-P. Wayne, PA: NCCLS; 1990. Iron Panel of the International Committee for Standardization in Haematology. Revised Recommendations for the Measurements of Serum Iron In Human Blood. Br J Haematology. 75:615-616; 1990.

NCCLS. Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition. NCCLS Document C24. Wayne, PA: NCCLS; 1999. 10. NCCLS. Interference Testing in Clinical Chemistry. NCCLS Document EP7. Wayne, PA: NCCLS; 1986. 11. Young DS. Effects of Drugs on Clinical Laboratory Tests, ed. 4. Washington D.C.: AACC Press; 1995. 12. Friedman RB, Young DS. Effects of Disease on Clinical Laboratory Tests. Washington, D.C.: AACC Press; 1990. 13. NCCLS. Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline. NCCLS Document EP9. Wayne, PA: NCCLS; 1995. 14. NCCLS. User Evaluation of Precision Performance with Clinical Chemistry Devices. NCCLS Document EP5. Wayne, PA: NCCLS; 1992.

Glossary of Symbols
The Glossary of Symbols table defines the symbols used on the VITROS Chemistry Products packaging and on the VITROS Chemistry Products Assay Sheets.

Glossary of Symbols
Do Not Reuse Use by or Expiration Date (YYYY-MM-DD) Lot Number Manufacturer's Serial Number Catalog Number or Product Code Attention: See Instructions for Use. JT Jf ^1 | EC [ HEP j S, v'7 V Manufacturer Authorized Representative Contains Sufficient for "n" Tests For In Vitro Diagnostic Use Store At or Below Store At or Above Store Between Consult Instructions for Use Fragile, Handle with Care. Keep Dry This end up

Version 1.0

Pub. No. C-371

VI"TFJCp*S 0

FeDT
Iron

INSTRUCTIONS FOR USE


Revision History

Revision History
Date of Revision 2003-10-01 Version 1.0 Description of Technical Changes* New format New organization and sections consistent with IVD Directive > Quality Control Material Selection - added statement regarding ethylene glycol > Known Interferences - removed statement regarding cupramine; updated bias value for imferon > Method Comparison - updated all comparisons and plots Precision - updated all data > References - added all except 4,11,12

The change bars indicate the position of a technical amendment to the text with respect to the previous version of the document.

When this Instructions For Use is replaced, sign and date below and retain as specified by local regulations or laboratory policies, as appropriate.

Signature

Obsolete Date

C
REP

Ortho-Clinical Diagnostics Mandeville House 62 The Broadway Amersham Buckinghamshire HP7 OHJ England Ortho-Clinical Diagnostics, Inc. 100 Indigo Creek Drive Rochester, NY 14626-5101

' Ortho-Clinical Diagnostics


company

Mil

VITROS is a trademark of Ortho-Clinical Diagnostics, Inc. Ortho-Clinical Diagnostics, Inc., 2003. All rights reserved. Printed in USA.

Pub. No. C-371

Version 1.0

Produ cts

VITR
Chemistry

INSTRUCTIONS FOR USE


VITROS Chemistry Products GGT DT Slides Intended Use

GGTDT
Gamma Glutamyltransferase

For in vitro diagnostic use only. VITROS GGT DT Slides quantitatively measure gamma glutamyltransferase (GGT) activity in serum and plasma.

Summary and Explanation of the Test


y-Glutamyltransferase plays a major role in glutathione metabolism and in resorption of amino acids from the glomerular filtrate. It is also important in the absorption of amino acids from the intestinal lumen. GGT is found mainly in the liver, pancreas, and kidney, although lower activities can be demonstrated in most other organs. Serum GGT is a sensitive indicator of hepatobiliary disease and is useful in the diagnosis of obstructive jaundice and chronic alcoholic liver disease, in the follow-up of chronic alcoholics undergoing treatment, and in the detection of hepatotoxicity. GGT is more responsive to biliary obstruction than AST, ALT, or ALKP. GGT is also increased in hepatoma, carcinoma of the pancreas, and carcinoma metastatic to the liver.1

Principles of the Procedure


The VITROS GGT DT Slide method is performed using the VITROS GGT DT Slide.and the VITROS Chemistry Products DT Calibrator Kit on VITROS DT60/DT60 II Chemistry Systems. The VITROS GGT DT Slide is a multilayered, analytical element coated on a polyester support. A drop of patient sample is deposited on the slide and is evenly distributed by the spreading layer to the underlying layers. GGT catalyzes the transfer of the y-glutamyl portion of L-y-glutamyl-p-nitroanilide to glycylglycine, simultaneously producing p-nitroaniline. The rate of change in reflection density is measured and is used to calculate the enzyme activity of GGT.

Reaction Sequence
L-y-glutamyl-p-nitroanilide + glycylglycine p-nitroaniline + y-glutamyl glycylglycine

Test Type and Conditions


Test Type and Conditions for GGT DT
Test Type Rate VITROS DT60/DT60 II Module DTSC/DTSC II Approximate Incubation Time 5 minutes Temperature 37C (98.6F) Wavelength 400 nm Sample Drop Volume 10 uL

Warnings and Precautions


For in vitro diagnostic use only. Take care when handling materials and samples of human origin. Since no test method can offer complete assurance that infectious agents are absent, consider all clinical specimens, controls, and calibrators potentially infectious. Handle specimens, solid and liquid waste, and test components in accordance with local regulations and NCCLS Guideline M292 or other published biohazard safety guidelines. For specific warnings and precautions for calibrators, quality control materials, and other components, refer to the instructions for use for the appropriate VITROS product, or to other manufacturer's product literature.

Version 1.0

Pub. No. C-343

GGTDT
Gamma Glutamyltransferase

INSTRUCTIONS FOR USE


Reagents

Reagents
Slide Diagram

Slide Ingredients
Reactive ingredients per cm Glycylglycine 0.2 mg; and L-y-glutamyl-p-nitroanilide 16 ug. Other ingredients Polymer beads, binders and surfactants.
\r

.
2

t *
3

1. Upper slide mount 2. Spreading layer (beads) 3. Reagent layer


buffer, pH 8.0 glycylglycine L-7-glytamyl-p-nitroanilide

-"" '_,

4. Support layer Lower slide mount


^ - 8

Slide Handling
Do not use slides with damaged or incompletely sealed packaging.

Slide Preparation
IMPORTANT: Theslide must reach room temperature, 18-28C (64-82F), before the wrapperis opened. Do not use unopened slides that have been at room temperature, 18-28C (64 "-82 F) for >48 hours. 1. Remove the unopened slide from the box. 2. Warm the unopened slide for 15 minutes. Unopened slides that remain in the sealed wrapper may be returned to refrigerator or freezer storage. 3. Remove the packaging and place the slide into the loading station within 15 minutes after opening.

Slide Storage and Stability


VITROS GGT DT Slides are stable until the expiration date on the carton when they are stored and handled as specified. Slide Storage and Stability for GGT DT Storage Condition Slides Room temperature 18-28C (64-82F) Unopened Refrigerated 2-8C (36-46F) Frozen <-18C(<0F) Room temperature 18-28C (64-82F) Opened

Stability <48 hours Until expiration date Until expiration date <15 minutes

Specimen Requirements
WARNING: Handle specimens as biohazardous material.

Specimens Recommended
Serum Plasma: EDTA Heparin Certain collection devices have been reported to affect other analytes and tests3 Confirm that your collection devices are compatible with this test.

Serum and Plasma Specimen Collection and Preparation


Collect specimens using standard laboratory procedures.4 6 Patient Preparation No special patient preparation is necessary. Special Precautions For the affect of high concentration of bilirubin on test results, refer to "Limitations of the Procedure." Centrifuge specimens and remove the serum or plasma from the cellular material within 4 hours of collection.6

Specimen Handling and Storage


Pub. No. C-343 Version 1.0

| i | V1TRI

INSTRUCTIONS FOR USE


Testing Procedure
Handle specimens as biohazardous material. Handle and store specimens in stoppered containers to avoid contamination and evaporation. Mix samples by gentle inversion and bring to room temperature, 18-28C (64-82F), prior to analysis.

GGT DT
Gamma Glutamyltransferase

Specimen Storage and Stability for GGT DT: Serum and Plasma6
Storage Room temperature Refrigerated Frozen Temperature 18-28C (64-82F) 2-8C (36-46F) <-18C(<0F) Stability <7 days <7 days <2 months

Testing Procedure
Materials Provided
VITROS Chemistry Products GGT DT Slides

Materials Required But Not Provided


. VITROS Chemistry Products DT Calibrator Kit Quality control materials, such as VITROS Chemistry Products DT Control I & II VITROS Chemistry Products 7% BSA Isotonic saline VITROS DT Pipette

Operating Instructions
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System. Bring all fluids and samples to room temperature, 18-28C (64-82F), prior to analysis. IMPORTANT;

Sample Dilution
| If gamma glutamyltransferase concentrations exceed the system's reportable (dynamic) range: 1. Dilute the sample with VITROS 7% BSA or isotonic saline. 2. Reanalyze. 3. Multiply the results by the dilution factor to obtain an estimate of the original sample's gamma glutamyltransferase activity.

Calibration
Required Calibrators
VITROS Chemistry Products DT Calibrator Kit, bottles 1, 2, and 4

Calibrator Preparation, Handling, and Storage


Refer to the instructions for use for VITROS DT Calibrator Kit.

Calibration Procedure
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

When to Calibrate
Calibrate: When the slide lot number changes. When critical system parts are replaced due to service or maintenance. When government regulations require. - For example, in the USA, CLIA regulations require calibration or calibration verification at least once every six months. The VITROS GGT DT test may also need to be calibrated: If quality control results are consistently outside acceptable range. After certain service procedures have been performed. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Version 1.0

Pub. No. C-343

GGTDT
Gamma Glutamyltransferase

INSTRUCTIONS FOR USE


Quality Control

Calculations
Based on sequential readings of the slide's reflectance at 400 nm over the defined incubation period, a rate of change in reflectance is determined. This rate is used in the software-resident multi-point rate calibration model to compute enzyme activity. Once a calibration has been performed for each slide lot, gamma-glutamyltransferase activity in unknown samples can be determined from the rate of change in reflectance measured for each unknown test slide.

Validity of a Calibration
Calibration parameters are automatically assessed by the VITROS DT60/DT60II Chemistry System against a set of quality parameters resident within the analyzer software. Failure to meet any of the pre-defined quality parameters results in a failed calibration. The calibration report should be used in conjunction with quality control results to determine the validity of a calibration.

Reportable (Dynamic) Range Reportable (Dynamic) Range for GGT DT


Conventional and SI Units (U/L)

5-1400
For out-of-range samples, refer to "Sample Dilution."

Traceability of the Calibration


I I Values assigned to the VITROSChemistry Products DT Calibrator Kit for gamma-glutamyltransferase are traceable to the gamma-glutamyltransferase method recommended by the International Federation of Clinical Chemistry (IFCC),7 adapted to a centrifugal analyzer at 37C.

Quality Control
Procedure Recommendations
| WA R N l N G: Handle quality control materials as biohazardous material. Choose control levels that check the clinically relevant range. Analyze quality control materials in the same manner as patient samples, before or during patient sample processing. To verify system performance, analyze control materials: - After calibration. - According to local regulations or at least once each day that the test is being performed. - After specified service procedures are performed. Refer to the operator's manual for your VITROS DT60/DT60 II System. If control results fall outside your acceptable range, investigate the cause before deciding whether to report patient results. For general quality control recommendations, refer to Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition8 or other published guidelines. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Quality Control Material Selection


I IMPORTANT: VITROS DT Control I & II are recommended for use with the VITROS DT60/DT60 II Chemistry System. Evaluate the performance of other commercial control fluids for compatibility with this test before using for quality control.

Control materials other than VITROS DT Controls I & II may show a difference when compared with other gamma glutamyltransferase methods if they: - Depart from a true human matrix. - Contain high concentrations of preservatives, stabilizers, or other nonphysiological additives. Enzyme activity might also vary with enzyme source, diluent temperature, and activation time during reconstitution. Do not use control materials stabilized with ethylene glycol.

Quality Control Material Preparation and Storage


Refer to the instructions for use for VITROS Chemistry Products DT Control I & II or to other manufacturer's product literature.

Pub. No. C-343

Version 1.0

INSTRUCTIONS FOR USE


Expected Values and Reporting Units

GGTDT
Gamma Glutamyltransferase

Expected Values and Reporting Units


Reference Interval
These reference intervals are the central 95% of results from an external study of 493 apparently healthy adult volunteers (277 females and 216 males).9 The study was conducted following NCCLS Protocol C28.10

Reference Interval for GGT DT


Conventional and SI Units (U/L) Adult Females Males 12-58 12-43 15-73

Each laboratory should confirm the validity of these intervals for the population it serves.

Reporting Units and Unit Conversion


The VITROS DT60/DT60 II Chemistry System may be programmed to report gamma glutamyltransferase results in conventional and SI units.

Reporting Units for GGT DT Conventional and SI Units


U/L

Limitations of the Procedure


Known Interferences
The VITROS GGT DT Slide method was screened for interfering substances following NCCLS Protocol EP7.11 The substances listed in the table, when tested at the concentrations indicated, caused the bias shown.

Known Interfering Substances for GGT DT


Gamma Glutamyltransferase Activity Conv./SI Units (U/L) 190 1100

Interferent*
Bilirubin**
*

Interferent Concentration 40 mg/dL 40 mg/dL (684 umol/L) (684 umol/L)

Average Bias Conv./SI Units (U/L) +6 -460

**

It is possible that other interfering substances may be encountered. These results are representative; however, your results may differ somewhat due to test-to-test variation. The degree of interference at concentrations other than those listed might not be predictable. Grossly elevated concentrations

Other Limitations
Certain drugs and clinical conditions are known to alter gamma glutamyltransferase activity in vivo. For additional information, refer to one of the published summaries.12 13

Version 1.0

Pub. No. C-343

GGTDT
Gamma Glutamyltransferase

INSTRUCTIONS FOR USE


Performance Characteristics

Performance Characteristics
Method Comparison

The plot and table show the results of a comparison of samples analyzed on the VITROS DT60 II System with those analyzed using the VITROS 950 System. Testing followed NCCLS Protocol EP9.14

Method Comparison for GGT DT: Serum


Conventional and SI Units 1500 1 y =x

1200 " 900 " 600 o 300 " 0 300 600 900 1200 1500
Comparative Method: VITROS 950 Systam (U/L)

Method Comparison for GGT DT: Serum


Conventional and SI Units (U/L) Correlation Range of Slope Coefficient Sample Activity Intercept Sy.x 1.00 0.999

n
DT60 II System vs. 950 System 60

18-1296

4.63

20.83

Precision
Precision was evaluated with quality control materials on the VITROS DT60 II System following NCCLS Protocol EP5.15 The data presented are a representation of test performance and are provided as a guideline. Variables such as sample handling and storage, reagent handling and storage, laboratory environment, and system maintenance can affect reproducibility of test results.

Precision for GGT DT: Serum


Conventional and SI Units (U/L) System VITROS DT60 II Mean Activity 77 417 Within Day SD* 0.5 3.4 Within Lab SD** Within Lab CV%** 1.0 4.7 1.3 1.1 No Observ. 88 87 No. Days
22

22

Within Day precision was determined using two runs/day with two replications. Within Lab precision was determined using a single lot of slides and calibrating weekly.

Pub. No. C-343

Version 1.0

INSTRUCTIONS FOR USE


References

GCTDT
Gamma Glutamyltransferase

References
Tietz NW(ed). Fundamentals of Clinical Chemistry, ed. 3. Philadelphia: WB Saunders; 391-392; 1987. NCCLS. Protection of Laboratory Workers from Instrument Biohazards and Infectious Diseases Transmitted by Blood, Body Fluids and Tissue; Approved Guideline. NCCLS Document M29 (OSBN 1-56238). NCCLS, Wayne, PA 19087; 1997. 3. Calam RR. Specimen Processing Separator Gels: An Update. J. Clin. Immunoassay. 11:86-90; 1988. 4. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Venipuncture. NCCLS Document H3. Wayne, PA: NCCLS; 1991. 5. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Skin Puncture. NCCLS Document H4. Wayne, PA: NCCLS; 1991. 6. Clinical Laboratory Handbook for Patient Preparation and Specimen Handling. Fascicle VI: Chemistry/Clinical Microscopy. Northfield, IL: College of American Pathologists; 1992. 7. Shaw L M, Stromme J H, London JL, Theodorsen L. IFCC methods for the measurement of catalytic concentration of enzymes. Part 4. IFCC method forgamma-glutamyltransferase. J, Clin. Chem. Clin. Biochem. 21: 633-646; 1983. 8. NCCLS. Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition. NCCLS Document C24. Wayne, PA: NCCLS; 1999. 9. Neels H, Van Boeckel E, Wauters A. Algemeen Ziekenhuis Middelheim, 1998. Data on file. 10. NCCLS. How to Define and Determine Reference Intervals in the Clinical Laboratory; Approved Guideline. NCCLS document C28-A (ISBN 1-56238-269-1). NCCLS, 940 West Valley Road, Suite 1400, Wayne, Pennsylvania 19087, 1995. 11. NCCLS. Interference Testing in Clinical Chemistry. NCCLS Document EP7. Wayne, PA: NCCLS; 1986. 12. Young DS. Effects of Drugs on Clinical Laboratory Tests, ed. 4. Washington D.C.: AACC Press; 1995. 13. Friedman RB, Young DS. Effects of Disease on Clinical Laboratory Tests. Washington, D.C.: AACC Press; 1990. 14. NCCLS. Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline. NCCLS Document EP9. Wayne, PA: NCCLS; 1995. 15. NCCLS. User Evaluation of Precision Performance with Clinical Chemistry Devices. NCCLS Document EP5. Wayne, PA: NCCLS; 1992. 1. 2.

Glossary of Symbols
The Glossary of Symbols table defines the symbols used on the VITROS Chemistry Products packaging and on the VITROS Chemistry Products Assay Sheets. Glossary of Symbols
Do Not Reuse Use by or Expiration Date (YYYY-MM-DD) Lot Number Manufacturer Authorized Representative Contains Sufficient for "n" Tests For In Vitro Diagnostic Use Store At or Below Store At or Above Store Between Consult Instructions for Use Fragile, Handle with Care. Keep Dry This end up

SN
REF

Manufacturer's Serial Number Catalog Number or Product Code Attention: See Instructions for Use.

Version 1.0

Pub. No. C-343

GGTDT
Gamma Glutamyltransferase

INSTRUCTIONS FOR USE


Revision History

Revision History
Date of Revision 2003-08-11 Version 1.0 Description of Technical Changes* > New format New organization and sections consistent with IVD Directive Specimen Collection and Preparation - removed statement regarding hemolyzed specimens. > Materials Required But Not Provided - added VITROS 7% BSA Reference Interval - updated all data > Limitations of the Procedure - added the Known Interfering Substances table Method Comparison - updated comparisons and the plot > Precision - updated all values > References - added all except 9, 10

The change bars indicate the position of a technical amendment to the text with respect to the previous version of the document.

When this Instructions For Use is replaced, sign and date below and retain as specified by local regulations or laboratory policies, as appropriate.

Signature

Obsolete Date

C
Ortho-Clinical Diagnostics Mandeville House 62 The Broadway Amersham Buckinghamshire HP7 OHJ England Ortho-Clinical Diagnostics, Inc. 100 Indigo Creek Drive Rochester, NY 14626-5101

'Ortho-Clinical Diagnostics
<4^ofMMeH company

VITROS is a trademark of Ortho-Clinical Diagnostics, Inc. Ortho-Clinical Diagnostics, Inc., 2003. All rights reserved. Printed in USA.

Pub. No. C-343

Version 1.0

I P roducts

VITRmB
Chemistry!

INSTRUCTIONS fOR USE


VITROS Chemistry Products GLU DT Slides Intended Use
For in vitro diagnostic use only. VITROS GLU DT Slides quantitatively measure glucose (GLU) concentration in serum and plasma.

GLUDT
Glucose

Summary and Explanation of the Test


Glucose is a primary cellular energy source. Fasting plasma glucose concentrations and tolerance to a dose of glucose are used to establish the diagnosis of diabetes mellitus and disorders of carbohydrate metabolism. Glucose measurements are used to monitor therapy in diabetics and in patients with dehydration, coma, hypoglycemia, insulinoma, acidosis, and ketoacidosis.1

Principles of the Procedure


The VITROS GLU DT Slide method is performed using the VITROS GLU DT Slide and the VITROS Chemistry Products DT Calibrator Kit on VITROS DT60/DT60 II Chemistry Systems. The VITROS GLU DT Slide is a multilayered, analytical element coated on a polyester support. A drop of patient sample is deposited on the slide and is evenly distributed by the spreading layer to the underlying layers. The oxidation of sample glucose is catalyzed by glucose oxidase to form hydrogen peroxide and gluconate. This reaction is followed by an oxidative coupling catalyzed by peroxidase in the presence of dye precursors to produce a dye. The intensity of the dye is measured by reflected light. The dye system used is closely related to that first reported by Trinder.2The chemistry of the glucose slides has been described by Curme, et al.3

Reaction Sequence
p-D-glucose + O2 + H2O
glucose oxidase

D-gluconic acid + H2O2


peroxidase

2H2O2 + 4-aminoantipyrine + 1,7-dihydroxynaphthalene

->

red dye

Test Type and Conditions


Test Type and Conditions for GLU DT
Test Type Colorimetric VITROS DT60/DT60 II Module DT60/DT60 II Approximate Incubation Time 5 minutes Temperature 37C (98.6F) Wavelength 555 nm Sample Drop Volume 10 |jL

Warnings and Precautions


For in vitro diagnostic use only. Take care when handling materials and samples of human origin. Since no test method can offer complete assurance that infectious agents are absent, consider all clinical specimens, controls, and calibrators potentially infectious. Handle specimens, solid and liquid waste, and test components in accordance with local regulations and NCCLS Guideline M294 or other published biohazard safety guidelines. For specific warnings and precautions for calibrators, quality control materials, and other components, refer to the instructions for use for the appropriate VITROS product, or to other manufacturer's product literature.

Version 1.0

Pub. No. C-300

VITR

GLUDT
Glucose

INSTRUCTIONS FOR USE


Reagents

Reagents
Slide Diagram

Slide Ingredients
Reactive ingredients per cm
2

__

' t

Glucose oxidase (Aspergillus Niger, E.C.1.1.3.4) 0.77 U; peroxidase (horseradish root, E.C.1.11.1.7) 3.6 U; 1,7-dihydroxynaphthalene (dye precursor) 67 \ig and 4-aminoantipyrine hydrochloride (dye precursor) 0.11 mg.
*~ -4

Other ingredients
Pigment, binders, buffer, surfactants, stabilizers and cross-linking agent.

1. Upper slide mount 2 Spreading layer (TIO2) Reagent layer glucose oxidase peroxidase dye precursors buffer, pH 5.0 4. Support layer S. Lower slide mount

Slide Handling
CAUTION: Do not use slides with damaged or incompletely sealed packaging.

Slide Preparation
IMPORTANT: The slide must reach room temperature, 18 "-2B <C (64 "-82 F), before the wrapper is opened. Do not use unopened slides that have been at room temperature, 18-28X! (64 "-82 F) for >48 hours. 1. 2. Remove the unopened slide from the box. Warm the unopened slide for 15 minutes. Unopened slides that remain in the sealed wrapper may be returned to refrigerator or freezer storage. 3. Remove the packaging and place the slide into the loading station within 15 minutes after opening.

Slide Storage and Stability


VITROS GLU DT Slides are stable until the expiration date on the carton when they are stored and handled as specified.

Slide Storage and Stability for GLU DT


Slides Unopened* Specimen Type Used All recommended specimens Plasma (Sodium fluoride/ potassium oxalate) Serum, Plasma (EDTA, Heparin) All recommended specimens Frozen Refrigerated Refrigerated Room temperature Storage Condition <-18C (<0F) 2-8C (36-46F) 2-8C (36-46F) 18-28C(64-82F) Stability Until expiration date <4 months Until expiration date <15 minutes

Opened

Do not store with or near hydrogen peroxide.

Specimen Requirements
W&RN1NG: Handle specimens as biohazardous material.

Specimens Recommended
. I I Serum Plasma: EDTA Heparin Sodium fluoride/potassium oxalate (see the Slide Storage and Stability table for slide storage when using this specimen type) Certain collection devices have been reported to affect other analytes and tests5 Confirm that your collection devices are compatible with this test.

IMPORTANT:

Serum and Plasma


Specimen Collection and Preparation
Collect specimens using standard laboratory procedures.6 7

Pub. No. C-300

Version 1.0

VITF-jjnpS

INSTRUCTIONS FOR USE


Testing Procedure
Patient Preparation No special patient preparation is necessary.

GLURT
Glucose

Special Precautions For the effect of sample hemolysis on test results, refer to "Limitations of the Procedure." Grossly lipemic samples must be diluted prior to analysis. Refer to "Sample Dilution "for dilution instructions. For the effect of elevated lipids on test results, refer to "Limitations of the Procedure." Particulate matter (for example, fibrin) in sufficient quantity may coat the spreading layer and limit diffusion of oxygen, causing a negative interference. To minimize particulate matter, do not centrifuge specimens until clotting is complete. Serum: - Centrifuge specimen at 1000X g for 10 minutes and remove serum from the clot within 30 minutes after collecting the specimen to avoid metabolism of glucose by the cells (approximately 7% per hour at room temperature).6 Heparin or EDTA plasma: - Follow manufacturer's recommendations for mixing anticoagulant with specimens. - Centrifuge specimen at 1000X g for 10 minutes and remove plasma from the cells within 30 minutes after collecting the specimen to avoid metabolism of glucose by the cells (approximately 7% per hour at room temperature).6 Sodium fluoride/potassium oxalate plasma: - Follow manufacturer's recommendations for mixing anticoagulant with specimens. - Centrifuge specimens and remove the plasma from the cells within 24 hours of collection.8 IMPORTANT: See the Slide Storage and Stability table for slide storage when using sodium fluoride/potassium oxalate plasma.

Specimen Handling and Storage " *j Handle specimens as biohazardous material. Handle and store specimens in stoppered containers to avoid contamination and evaporation. Mix samples by gentle inversion and bring to room temperature, 18-28C (64-82F), prior to analysis. Specimen Storage and Stability for GLU DT: Serum and Plasma8 Temperature Stability Storage 18-28OC(64-82F) Room temperature <24 hours Refrigerated 2-8C (36-46F) <7 days Frozen <-18C(<0F) <1 year

Testing Procedure
Materials Provided
VITROS Chemistry Products GLU DT Slides

Materials Required But Not Provided


VITROS Chemistry Products DT Calibrator Kit Quality control materials, such as VITROS Chemistry Products DT Control I & II Isotonic saline Reagent-grade water VITROS DT Pipette

Operating Instructions
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System. Bring all fluids and samples to room temperature, 18-28C (64-82F), prior to analysis. IMPORTANT:

Sample Dilution
If glucose concentrations exceed the system's reportable (dynamic) range: 1. Dilute the sample with isotonic saline or reagent-grade water. 2. Reanalyze. 3. Multiply the results by the dilution factor to obtain an estimate of the original sample's glucose concentration.

Version 1.0

Pub. No. C-300

GLUDT
Glucose

INSTRUCTIONS FOR USE


Calibration

Calibration
Required Calibrators
VITROS Chemistry Products DT Calibrator Kit, bottles 1, 2, and 3

Calibrator Preparation, Handling, and Storage


Refer to the instructions for use for VITROS DT Calibrator Kit.

Calibration Procedure
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

When to Calibrate
Calibrate: When the slide lot number changes. When critical system parts are replaced due to service or maintenance. When government regulations require. - For example, in the USA, CLIA regulations require calibration or calibration verification at least once every six months. The VITROS GLU DT test may also need to be calibrated: If quality control results are consistently outside acceptable range. After certain service procedures have been performed. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Calculations

Reflectance from the slide is measured at 555 nm after the fixed incubation time. Once a calibration has been performed for each slide lot, glucose concentration in unknown samples can be determined using the software-resident endpoint colorimetric math model and the response obtained from each unknown test slide.

Validity of a Calibration
Calibration parameters are automatically assessed by the VITROS DT60/DT60 II Chemistry System against a set of quality parameters resident within the analyzer software. Failure to meet any of the pre-defined quality parameters results in a failed calibration. The calibration report should be used in conjunction with quality control results to determine the validity of a calibration.

Reportable (Dynamic) Range Reportable (Dynamic) Range for GLU DT


Conventional (mg/dL) SI Units (mmol/L) 1.1-25.0

20-450

For out-of-range samples, refer to "Sample Dilution."

Traceability of the Calibration


Values assigned to the VITROS Chemistry Products DT Calibrator Kit for glucose are traceable to the Certified NIST (National Institute of Standards and Technology) Reference Material, SRM (Standard Reference Material) 917b. The Ortho-Clinical Diagnostics calibration laboratory uses SRM 917b to calibrate the CDC Hexokinase method9 to support glucose value assignment for the VITROS DT Calibrator Kit.

Quality Control
Procedure Recommendations
| >' ' Handle quality control materials as biohazardous material. Choose control levels that check the clinically relevant range. Analyze quality control materials in the same manner as patient samples, before or during patient sample processing. To verify system performance, analyze control materials: - After calibration. - According to local regulations or at least once each day that the test is being performed. - After specified service procedures are performed. Refer to the operator's manual for your VITROS DT60/DT60 II System. If control results fall outside your acceptable range, investigate the cause before deciding whether to report patient results.

Pub.

No. C-300

Version 1.0

INSTRUCTIONS FOR USE


Expected Values and Reporting Units

GLUDT
Glucose

For general quality control recommendations, refer to Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Bditionm or other published guidelines. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Quality Control Material Selection


IMPORTANT: VITROS DT Control I & II are recommended for use with the VITROS DT60/DT60 II Chemistry System. Evaluate the performance of other commercial control fluids for compatibility with this test before using for quality control.

Control materials other than VITROS DT Controls I & II may show a difference when compared with other glucose methods if they: - Depart from a true human matrix. - Contain high concentrations of preservatives, stabilizers, or other nonphysiological additives. Do not use control materials stabilized with ethylene glycol.

Quality Control Material Preparation and Storage


Refer to the instructions for use for VITROS Chemistry Products DT Control I & II or to other manufacturer's product literature.

Expected Values and Reporting Units


Reference Interval
These reference intervals are based on external studies for serum.1 Reference Interval for GLU DT Conventional Units (mg/dL) Fasting adults 74-106

SI Units (mmol/L) 4.1-5.9

Each laboratory should confirm the validity of these intervals for the population it serves.

Reporting Units and Unit Conversion


The VITROS DT60/DT60 II Chemistry System may be programmed to report glucose results in conventional and SI units. Reporting Units and Unit Conversion for GLU DT Conventional Units SI Units mg/dL mmol/L (mg/dL x 0.05551)

Limitations of the Procedure


Known Interferences
In fresh specimens, catalase released from the lysis of red blood cells causes a negative bias in glucose results. The degree of bias is proportional to the degree of hemolysis. In fresh samples, a negative bias of up to 10% may be observed with a level of hemolysis associated with a hemoglobin concentration of 250 mg/dL (2.5 g/L). Catalase activity decreases with sample storage. Aged samples that are hemolyzed may exhibit a positive bias of up to 10% due to the spectral interference of hemoglobin. Therefore, the magnitude and direction of bias observed with hemolyzed specimens will vary due to the level of catalase activity and concentration of hemoglobin present in the sample.

NOTE:

Elevated lipids may limit diffusion of oxygen to the reactants. Dilute grossly lipemic samples before analysis. The VITROS GLU DT Slide method was screened for interfering substances following NCCLS Protocol EP7.12 The substances listed in the table, when tested at the concentrations indicated, caused the bias shown.

Known Interfering Substances for GLU DT


Interfered* Interferent Concentration 5 g/dL 10g/dL (50 g/L) (100 g/L) Glucose Concentration Conv. (mg/dL) SI (mmol/L) 100 5.55 100 5.55 Average Bias Conv. (mg/dL) SI (mmol/L) -5 -0.28 +6 +0.33

It is possible that other interfering substances may be encountered. These results are representative; however, your results may differ somewhat due to test-to-test variation. The degree of interference at concentrations other than those listed might not be predictable.

Version 1.0

Pub. No, C-300

GIUDT
Glucose

INSTRUCTIONS FOR USE


Performance Characteristics Certain drugs and clinical conditions are known to alter glucose concentrations in vivo. For additional information, refer to one of the published summaries. 13 ' u

Other Limitations

Performance Characteristics
Method Comparison

The plots and table show the results of a comparison of samples analyzed on the VITROS DT60 II System with those analyzed using the VITROS 950 System. Testing followed NCCLS Protocol EP9.15 Method Comparison for G L U DT: Serum Conventional Units 500 450 * 400 30 SI Units y =x

g-

I I a ft

350 " 300 " 250 '

a>

20 15 10 5 0

I
o | 5

2()

<> "

150 " 100 0 50 100 200 300 400 500


Comparative Method: VITROS 950 System (mg/dL)

10

15

20

25

30

Comparative Method: VITROS 950 System (mmol/L)

Method Comparison for GLU DT: Serum


Conventional Units (mg/dL) n DT60 II System vs. 950 System 55 Correlation Slope Coefficient 1.02 1.000 Range of Sample Cone. 25-445 Intercept 0.67 Sy.x 3.78 SI Units (mmol/L) Range of Sample Cone. Intercept 1.4-24.7 0.04

Sy.x 0.21

Precision
Precision was evaluated with quality control materials on the VITROS DT60 II System following NCCLS Protocol EP5.' 6 The data presented are a representation of test performance and are provided as a guideline. Variables such as sample handling and storage, reagent handling and storage, laboratory environment, and system maintenance can affect reproducibility of test results.

Precision for GLU DT: Serum


Conventional Units (mg/dL) System VITROS DT60 II Mean Cone. 91 309 Within Day SD* 1.0 3.8 Within Lab SD** 1.7 4.4 SI Units (mmol/L) Mean Within Within Cone. Day SD* Lab SD** 5.0 0.05 0.09 17.2 0.25 0.21 Within Lab CV%** 1.8 1.4 No. Observ. 88 88 No. Days 22 22

Within Day precision was determined using two runs/day with two replications. Within Lab precision was determined using a single lot of slides and calibrating weekly.

Pub. No. C-300

Version 1.0

INSTRUCTIONS FOR USE


References

GLUDT
Glucose

References
1. 2. 3. 4. 5. 6. 7. 8. Tietz NW(ed). Textbook of Clinical Chemistry, ed. 2. Philadelphia: WB Saunders; 928-960; 1994. Trinder P. Determination of Glucose in Blood Using Glucose Oxidase with an Alternative Oxygen Receptor. Ann. Clin. Biochem. 6:24; 1969. Curme HG, et al. Multilayer Film Elements for Clinical Analysis. Clin. Chem. 24:1335-1342; 1978. NCCLS. Protection of Laboratory Workers from Instrument Biohazards and Infectious Diseases Transmitted by Blood, Body Fluids and Tissue; Approved Guideline. NCCLS Document M29 (OSBN 1-56238). NCCLS, Wayne, PA 19087; 1997. Calam RR. Specimen Processing Separator Gels: An Update. J Clin Immunoassay. 11:86-90; 1988. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Venipuncture. NCCLS Document H3. Wayne, PA: NCCLS;

1991.
NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Skin Puncture. NCCLS Document H4. Wayne, PA: NCCLS;

1991.
Clinical Laboratory Handbook for Patient Preparation and Specimen Handling. Fascicle VI: Chemistry/Clinical Microscopy. Northfield, IL: College of American Pathologists; 1992. 9. Neese JW, Duncan P, Bayse DD, et al. Development and Evaluation of a Hexokinase/Glucose-6-phosphate Dehydrogenase Procedure for Use as a National Glucose Reference Method. HEW Publication No. (CDC) 77-8330. HEW. USPHS, Centers for Disease Control; 1976. 10. NCCLS. Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition. NCCLS Document C24. Wayne, PA: NCCLS; 1999. 11. Tietz NW (ed). Textbook of Clinical Chemistry, ed. 3. Philadelphia: WB Saunders; 1815; 1999. 12. NCCLS. Interference Testing in Clinical Chemistry. NCCLS Document EP7. Wayne, PA: NCCLS; 1986. 13. Young DS. Effects of Drugs on Clinical Laboratory Tests, ed. 4. Washington D.C.: AACC Press; 1995. 14. Friedman RB, Young DS. Effects of Disease on Clinical Laboratory Tests. Washington, D.C.: AACC Press; 1990. 15. NCCLS. Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline. NCCLS Document EP9. Wayne, PA: NCCLS; 1995. 16. NCCLS. User Evaluation of Precision Performance with Clinical Chemistry Devices. NCCLS Document EP5. Wayne, PA: NCCLS;

1992.

Glossary of Symbols
The Glossary of Symbols table defines the symbols used on the VITROS Chemistry Products packaging and on the VITROS Chemistry Products Assay Sheets.

Glossary of Symbols
Do Not Reuse Use by or Expiration Date (YYYY-MM-DD) Lot Number Manufacturer's Serial Number Catalog Number or Product Code Attention: See Instructions for Use. EC | REP I \ y / \/ Manufacturer Authorized Representative Contains Sufficient for "n" Tests For In Vitro Diagnostic Use Store At or Below Store At or Above Store Between Consult Instructions for Use Fragile, Handle with Care. Keep Dry This end up

Version 1.0

Pub. No. C-300

GLUDT
Glucose

INSTRUCTIONS FOR USE


Revision History

Revision History
Date of Revision 2003-08-11 Version 1.0 Description of Technical Changes* New format New organization and sections consistent with IVD Directive Slide Storage and Stability - added the Specimen Type Used column; updated storage values for opened cartridges Specimens Recommended - added sodium fluoride/potassium oxalate Reference Interval - updated data Limitations of the Procedure - deleted statement regarding ascorbic acid; updated data for hemolysis Method Comparison - updated all comparisons and plots Precision - updated values References - added all

The change bars indicate the position of a technical amendment to the text with respect to the previous version of the document.

When this Instructions For Use is replaced, sign and date below and retain as specified by local regulations or laboratory policies, as appropriate.

Signature

Obsolete Date

C
EC

Ortho-Clinical Diagnostics Mandeville House 62 The Broadway Amersham Buckinghamshire HP7 OHJ England Ortho-Clinical Diagnostics, Inc. 100 Indigo Creek Drive Rochester, NY 14626-5101

' Ortho-Clinical Diagnostics


a flohtMMjjefMM8H company

VITROS is a trademark of Ortho-Clinical Diagnostics, Inc. Ortho-Clinical Diagnostics, Inc., 2003. All rights reserved. Printed in USA.

Pub.

No. C-300

Version 1.0

IProducts

VITRQ35
Chemistry!

INSTRUCTIONS FOR USE


VITROS Chemistry Products DT Micro HDL Cholesterol Kit Intended Use

HDLC DT
Micro HDL Cholesterol

For in vitro diagnostic use only. For use in the quantitative measurement of HDL cholesterol (HDLC) concentration in serum and plasma.

Summary and Explanation of the Test


HDL cholesterol is used to evaluate the risk of developing coronary heart disease (CHD). The risk of CHD increases with lower HDL cholesterol concentrations.

Principles of the Procedure


VITROS DT Micro HDL Cholesterol Kit is used to prepare and test samples for high density lipoprotein (HDL) cholesterol on VITROS DT60/DT60 II Chemistry Systems. The specimens must initially be treated with a reagent to remove other lipoproteins which also contain cholesterol, including the very low density (VLDL) and low-density (LDL) classes. The amount of HDL cholesterol can then be determined using VITROS Chemistry Products HDLC DT Slides. HDL is separated by the precipitation of LDL and VLDL using dextran sulfate (MW 50,000) and magnesium chloride1 that is provided in the VITROS Chemistry Products DT Micro HDL Tube. The HDL lipoproteins remain in the liquid portion of the tube after centrifugation. This liquid portion is called the supernate and is the portion analyzed. The non-HDL fractions form a pellet on the bottom of the tube and are discarded. A drop of pretreated patient sample is deposited on the slide and is evenly distributed by the spreading layer to the underlying layers. The Triton X-100 (TX100) surfactant in the spreading layer aids in dissociating the cholesterol and cholesterol esters from lipoprotein complexes present in the sample. Hydrolysis of the cholesterol esters to cholesterol is catalyzed by cholesterol ester hydrolase. Free cholesterol is then oxidized in the presence of cholesterol oxidase to form cholestenone and hydrogen peroxide. Finally, hydrogen peroxide oxidizes a leuco dye in the presence of peroxidase to generate a colored dye. The density of dye formed is proportional to the HDL cholesterol concentration present in the pretreated sample and is measured by reflectance spectrophotometry.

Reaction Sequence
high density lipoproteins cholesterol esters + H2O cholesterol + O2 H2O2 + leuco dye
cholesterol ester hydrolase

->

cholesterol + cholesterol esters + proteins - > cholesterol + fatty acids - > cholest-4-en-3-one + H2O2

cholesterol oxidase peroxidase

- > dye + 2H2O

Test Type and Conditions


Test Type and Conditions for HDLC DT
Test Type Colorimetric VITROS DT60/DT60 1 1 Module DT60/DT60 II Approximate Incubation Time 5 minutes Temperature 37C (98.6F) Wavelength 660 nm Sample Drop Volume 10 uL

Version 2.0

Pub. No. C-354 EN

VITRCp5 0

HDLC DT
Micro HDL Cholesterol

INSTRUCTIONS FOR USE


Warnings and Precautions

Warnings and Precautions


For in vitro diagnostic use only. Take care when handling materials and samples of human origin. Since no test method can offer complete assurance that infectious agents are absent, consider all clinical specimens, controls, and calibrators potentially infectious. Handle specimens, solid and liquid waste, and test components in accordance with local regulations and NCCLS Guideline M292 or other published biohazard safety guidelines. While VITROS HDLC Sample Diluent product is bovine in origin, it should be handled using the same precautions as with any other blood or blood-derived product. For specific warnings and precautions for calibrators, quality control materials, and other components, refer to the Instructions for Use for the appropriate VITROS product, or to other manufacturer's product literature.

Personal Protection and Ventilation


Wear impervious gloves and proper protective clothing. Safety glasses are recommended. Good general ventilation in the work area is recommended. Minimize the potential for production of aerosols.

Accidental Spillage and Disposal


WAP VITROS DT Micro HDL Tubes contain sodium azide. Disposal of reagent into sinks with copper or lead plumbing should be followed with plenty of water to prevent formation of potentially explosive metallic azides.

Absorb spilled material in vermiculite or other suitable absorbents; sweep up and dispose of with clinical waste. Disinfect area with a 5.25% sodium hypochlorite solution (household bleach) diluted with 9 parts water and leave undisturbed for at least 30 minutes. Excess or unwanted materials and packaging for this product should be disposed of with other clinical waste. VITROS HDL Cholesterol Reagent contains sodium azide (0.01%; 0.01 g/dL). Disposal of reagent into sinks with copper or lead plumbing should be followed with copious volumes of water to prevent formation of potentially explosive metallic azides.

First Aid
WARNSNC

VITROS DT Micro HDL Tubes contain gentamicin sulfate and sodium azide. R22 - Harmful if swallowed.

Inhalation - Remove to fresh air. Seek medical advice. Skin - Wash skin after each contact with soap and plenty of water for at least 15 minutes. Seek medical attention if skin is cut or punctured. Eye - Immediately flush eyes, including under the eyelids, with plenty of water for at least 15 minutes and get medical attention. Ingestion - Drink 1 -2 glasses of water. Seek medical advice.

Transportation
This product is not classified as a dangerous substance for transportation purposes. However, package with adequate protection to prevent breakage and ship under refrigerated conditions.

Reagents
Slide Diagram

HDLC DT Slide Ingredients


Reactive ingredients per cm
2

1
2

Triton X-100 0.8 mg; cholesterol oxidase (Nocardia, E.C.1.1.3.6) 0.2 U; cholesterol ester hydrolase (Candida rugosa, E.C.3.1.1.13) 0.7 U; peroxidase (horseradish root, E.C.1.11.1.7) 5.3 U; and 2-(3,5-dimethoxy-4-hydroxyphenyl)4,5-bis-(4-dimethylaminophenyl) imidazole (leucodye) 0.2 mg.

Other ingredients
Pigment, binder, buffer, surfactants, stabilizers and cross-linking agent.

1 . Upper slide mount 2. Spreading layer (BaSCUl Triton X-100 cholesterol ester hydrolase cholesterol oxidase peroxidase leuco dye 3. Sublayer buffer, pH 6.25 4. Support layer 5. Lower slide mount

DT Micro HDL Tubes Reactive Ingredients


Dextran sulfate (MW50,000) 0.08 mg, magnesium chloride hexahydrate 0.7 mg

Other ingredients
sodium azide 0.22%, gentamicin sulfate <0.01%, dye and preservatives Version 2.0

Pub. No. C-354 EN

INSTRUCTIONS FOR USE


Specimen Requirements

HDLC DT
Micro HDL Cholesterol

HDLC Sample Diluent


Reactive Ingredients
Bovine serum albumin 7%

Other ingredients
Sodium azide 0.05%, inorganic salts and preservatives

Slide Handling
Do not use slides with damaged or incompletely sealed packaging.

Slide Preparation
IMPORTANT: The slide must reach room temperature, 18-28 C (64 -82 F), before the wrapper is opened. Do not use unopened slides that have been at room temperature, 18 -28 C (64 -82F) for >48 hours. 1. Remove the unopened slide from the box. 2. Warm the unopened slide for 15 minutes. Unopened slides that remain in the sealed wrapper may be returned to refrigerator or freezer storage. 3. Remove the packaging and place the slide into the loading station within 15 minutes after opening.

Kit Storage and Stability


VITROS DT Micro HDL Cholesterol Kit components are stable until the expiration date on the carton when they are stored and handled as specified.

Kit Storage and Stability for HDLC DT


Kit Component Slide Unopened Opened Tubes Refrigerated Frozen Room temperature Room temperature Refrigerated Frozen Sample Diluent* Refrigerated Frozen * Discard if the solution becomes cloudy or turbid. Verify performance with quality control materials: - If the system is turned off for more than 2 hours. 2-8C (36-46F) <-18C(<0F) Until expiration date Until expiration date 2-8C (36-46F) <-18C(<0F) 18-28OC(64-82F) 18O-28C(64O-82F) 2-8C (36-46F) <-18C(<0F) Until expiration date Until expiration date <48 hours <15 minutes Until expiration date Until expiration date Storage Condition Stability

Specimen Requirements
Handle specimens as biohazardous material.

Specimens Recommended
Serum Plasma:3 Heparin Certain collection devices have been reported to affect other analytes and tests." Confirm that your collection devices are compatible with this test.

Specimens Not Recommended


. Plasma:5 EDTA

Version 2.0

Pub. No. C-354 EN

VITH[fiS@

HDIC DT
Micro HDL Cholesterol Serum and Plasma Specimen Collection and Preparation
Collect specimens using standard laboratory procedures.8 7

INSTRUCTIONS FOR USE


Specimen Pretreatment and Testing Procedure

Patient Preparation No special patient preparation is necessary unless the HDL cholesterol test is part of a complete lipid profile. Then, a 12- to 14-hour fast is necessary.1 Special Precautions Centrifuge specimens and remove the serum from the cellular material within 3 hours of collection.8 If further processing is delayed, store in the refrigerator at 2-8C (36-46F). Specimen Handling and Storage ' ' Handle specimens as biohazardous material.

Handle and store specimens in stoppered containers to avoid contamination and evaporation. Mix samples by gentle inversion and bring to room temperature, 18-28C (64-82F), prior to analysis.

Specimen Storage and Stability for HDLC DT: Serum and Plasma8
Storage Original Specimen Refrigerated Frozen Pretreated Specimen (Supemate) Freezer * Temperature 2-8C (36-46F) <-18C (SOF) | Stability <3 days <4 weeks* <3 months <2 years

<-20C (<-4F) <-70C (<-94F) For longer storage, separate HDL fractions and freeze the supernate. Avoid repeated freeze-thaw cycles.

IMPORTANT:

Specimen Pretreatment and Testing Procedure


Materials Provided
25 VITROS Chemistry Products HDLC DT Slides 27 VITROS Chemistry Products DT Micro HDL Tubes VITROS Chemistry Products HDLC Sample Diluent

Materials Required But Not Provided


. VITROS Chemistry Products DT Calibrator Kit VITROS Chemistry Products DT Controls I and II VITROS DT Pipette Pipette capable of accurately delivering 50 uL of serum or plasma to a VITROS DT Micro HDL Tube. Centrifuge capable of generating forces of 1500 x g, or a microcentrifuge capable of generating forces of 12,600 x g. Micro capillary collection device large enough to obtain at least 50 uL of serum or plasma. A vortex mixer is recommended

Special Precautions
Use a vortex speed that will not cause the mixture to spill out of the sample tube.

Procedure
IMPORTANT; Do not pretreat calibrators.

Be sure to use components from the same kit lot number. IMPORTANT: 1. Allow at least 10 minutes for refrigerated VITROS DT Micro HDL Tubes and VITROS HDLC Sample Diluent to reach room temperature. If these materials are stored at -18C (0F), they will require a longer warm-up period (at least 30 minutes). Once opened, the VITROS Micro HDLC Sample Diluent should be stored tightly capped in the refrigerator between uses. 2. Pipette 50 uL of serum or heparin plasma into the VITROS DT Micro HDL Tube.

Pub. No. C-354 EN

Version 2.0

INSTRUCTIONS FOR USE


Calibration
3. Cap and mix thoroughly for 30 seconds - a vortex mixer is recommended. NOTE: The sample will become cloudy during mixing.

HDLCDT
Micro HDL Cholesterol

4. Let stand for a minimum of 5 minutes. 5. Centrifuge the VITROS DT Micro HDL Tube for 10 minutes at 1,500 x g. Alternatively, you may centrifuge the tube for a shorter duration using a microcentrifuge for 95 seconds at 12,600 x g. 6. Visually check supernates for clarity. The non-HDL fractions form a pellet on the bottom of the tube. Do not disturb the pellet. If the supernate is clear, transfer cleared supernate directly from the VITROS DT Micro HDL Tube to a sample cup for analysis on the DT analyzer. IMPORT ANT: Supernates should be removed from the pelleted precipitate as soon as possible following centrifugation. The supernates should be used within 15 minutes. If analysis on the VITROS DT60/DT60 II Chemistry System is to be delayed more than one hour, the supernate should be refrigerated. If analysis is delayed longer than 8 hours, refer to the "Specimen Handling and Storage" section.

7. Analyze treated sample as instructed in the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Operating Instructions
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System. Bring all fluids and samples to room temperature, 18-28C (64-82F); analyze immediately. IMPORTANT:

Sample Dilution
If HDL concentrations exceed the system's reportable (dynamic) range or if the supernatant is cloudy or has floating particles after pretreatment: 1. Dilute the untreated sample with an equal volume of VITROS HDLC Sample Diluent. 2. Mix gently by inverting several times. 3. Pipette 50 uL of the diluted specimen into a second VITROS DT Micro HDL Tube. 4. Repeat steps 1 through 6 of the procedure above. Remember to multiply the result by the dilution factor of 2.0. 5. Reanalyze. 6. Multiply the results by 2 to obtain an estimate of the original sample's HDL concentration. NOTE: Additional dilutions are not recommended.

Calibration
Required Calibrators
VITROS Chemistry Products DT Calibrator Kit, bottles 1, 2, and 4

Calibrator Preparation, Handling, and Storage


IMPORTANT: Do not pretreat calibrators. Refer to the instructions for use for VITROS DT Calibrator Kit.

Calibration Procedure
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

When to Calibrate
Calibrate: When the slide lot number changes. When critical system parts are replaced due to service or maintenance. When government regulations require. - For example, in the USA, CLIA regulations require calibration or calibration verification at least once every six months. The VITROS HDLC DT slides may also need to be calibrated: If quality control results are consistently outside acceptable range. After certain service procedures have been performed. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Version 2.0

Pub. No. C-354 EN

HDICDT
Micro HDL Cholesterol

INSTRUCTIONS FOR USE


Quality Control

Calculations
Reflectance from the slide is measured at 660 nm after the fixed incubation time. Once a calibration has been performed for each slide lot, high-density lipoprotein cholesterol concentration in unknown samples can be determined using the softwareresident endpoint colorimetric math model and the response obtained from each unknown test slide.

Validity of a Calibration
Calibration parameters are automatically assessed by the VITROS DT60/DT60 II Chemistry System against a set of quality parameters resident within the analyzer software. Failure to meet any of the pre-defined quality parameters results in a failed calibration. The calibration report should be used in conjunction with quality control results to determine the validity of a calibration.

Reportable (Dynamic) Range Reportable (Dynamic) Range for HDLC DT


Conventional (mg/dL) 1-110 SI Units (mmol/L) 0.03-2.84

For out-of-range samples, refer to "Sample Dilution."

Traceability of the Calibration


Values assigned to the VITROS Chemistry Products DT Calibrator Kit for High Density Lipoprotein-Cholesterol (HDLC) Reagent with VITROS HDLC DT slides are traceable to the Certified NIST (National Institute of Standards and Technology) Reference Material, SRM* (Standard Reference Material) 911b. The Ortho-Clinical Diagnostics Calibration Laboratory uses SRM 911b to assign values to secondary standards for calibration of an HDLC Selected Method. The Selected Method supports VITROS HDL Cholesterol reagent value assignment for the VITROS DT Calibrator Kit. The HDLC Selected Method includes (MW 50,000) Dextran Sulfate precipitation of non-HDL lipoproteins according to the recommendations of the Centers for Disease Control (CDC),10 followed by automated enzymatic determination of supernate (HDL fraction) cholesterol concentration.

Quality Control
IMPORTANT: Controls must be pretreated.

Procedure Recommendations
| .. Handle quality control materials as biohazardous material. Choose control levels that check the clinically relevant range. Analyze quality control materials in the same manner as patient samples, before or during patient sample processing. To verify system performance, analyze control materials: - After calibration. - According to local regulations or at least once each day that the test is being performed. - After specified service procedures are performed. Refer to the operator's manual for your VITROS DT60/DT60II System. If control results fall outside your acceptable range, investigate the cause before deciding whether to report patient results. For general quality control recommendations, refer to Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition'1'' or other published guidelines. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Quality Control Material Selection


IMPORTANT: VITROS DT Control I & II are recommended for use with the VITROS DT60/DT60II Chemistry System. Evaluate the performance of other commercial control fluids for compatibility with this test before using for quality control.

Control materials other than VITROS DT Controls I & II may show a difference when compared with other HDL cholesterol methods if they: - Depart from a true human matrix. - Contain high concentrations of preservatives, stabilizers, or other nonphysiological additives. Do not use control materials stabilized with ethylene glycol.

Quality Control Material Preparation and Storage


Refer to the instructions for use for VITROS Chemistry Products DT Control I & II or to other manufacturer's product literature.

Pub. No. C-354 EN

Version 2.0

INSTRUCTIONS FOR USE


Interpretation of Results and Expected Results

HDIC DT
Micro HDL Cholesterol

Interpretation of Results and Expected Results


Interpretation of Results
LDL cholesterol and VLDL cholesterol can be calculated from the results of the VITROS CHOL DT Slide, VITROS TRIG DT Slide, and the results for HDL cholesterol to provide complete lipoprotein profiles. LDL = CHOL-HDLC-VLDL VLDL = TRIG/5 for conventional units (mg/dL) VLDL = TRIG/2.2 for SI units (mmol/L) Calculation of LDL is not appropriate for samples with triglyceride concentrations greater than 400 mg/dL (4.57 mmol/L) or with samples from patients who have type III hyperlipoproteinemia (electrophoresis "broad beta" lipoprotein) present.12

Expected Results
These guidelines have been recommended by the National Institutes of Health.'3

Reference Interval for HDLC DT


Low High Conv. Units (mg/dL) <40 >60 SI Units (mmol/L) <1.0 >1.6

Each laboratory should confirm the validity of these intervals for the population it serves.

Reporting Units and Unit Conversion


The VITROS DT60/DT60 II Chemistry System may be programmed to report HDL cholesterol results in conventional and SI units.

Reporting Units and Unit Conversion for HDLC DT Conventional Units SI Units
mg/dL mmol/L (mg/dL x 0.02586)

Limitations of the Procedure


Known Interferences
The VITROS DT Micro HDL Cholesterol Kit method was screened for interfering substances following NCCLS Protocol EP7." The substances listed in the table, when tested at the concentrations indicated, caused the bias shown.

Known Interfering Substances for HDLC DT


Interferent Interferent* Ascorbic acid Dipyrone Dopamine N-acetylcysteine
* **

HDL Cholesterol Concentration Conv. (mg/dL) 40.9 40.5 44.6 40.2 SI (mmol/L) 1.06 1.05 1.15 1.04

Bias** Conv. (mg/dL) -7 -7 -9 -6 SI (mmol/L) -0.18 -0.18 -0.22 -0.14

Concentration Comments 3 mg/dL 170umol/L High Therapeutic 12 mg/dL 3.6 mmol/L High IV Drip 4 mg/dL 200 umol/L __ High IV Drip 10 mg/dL 0.61 mmol/L Oral Therapeutic

It is possible that other interfering substances may be encountered. These results are representative; however, your results may differ somewhat due to test-to-test variation. The degree of interference at concentrations other than those listed might not be predictable. The bias is an estimate of the maximum difference observed.

Other Limitations
Certain drugs and clinical conditions are known to alter HDL cholesterol concentration in vivo. For additional information, refer to one of the published summaries.' 516 "

Version 2.0

Pub. No. C-354 EN

HD1CDT
Micro HDL Cholesterol

INSTRUCTIONS FOR USE


Performance Characteristics

Performance Characteristics
Method Comparison
The plots and table show the results of a comparison of the VITROS HDL Cholesterol Method analyzed on the VITROS DT60 II Chemistry System using the Dextran Sulfate/Enzymatic10 comparative method using pretreated serum specimens.

Method Comparison for HDLC DT: Serum


Conventional Units y =x
100

SI Units y =x

3.0 2.5

80

2.0 1.5
40 20

1.0 0.5 0.0 0 20 40 60 80 100 0.0 0.5 1.0 1.5 2.0 2.5 3.0

Comparative Method: Dextran Sulfate/Enzymatic (mg/dL)

Comparative Method: Dextran Sulfate/Enzymatic (mmol/L)

Method Comparison for HDLC DT: Serum


Conventional Units (mg/dL) n DT60 II System vs. comparative method 43 Correlation Slope Coefficient 1.00 0.998 Range of Sample Cone. Intercept Sy.x 0.02 1.23 SI Units (mmol/L) Range of Sample Cone. 0.3-2.6 Intercept 0.00 Sy.x 0.03

12-101

Precision
Precision was evaluated with quality control materials on VITROS the DT60 II System following NCCLS Protocol EP5.18 The data presented are a representation of test performance and are provided as a guideline. Variables such as sample handling and storage, reagent handling and storage, laboratory environment, and system maintenance can affect reproducibility of test results.

Precision for HDLC DT: Serum


Conventional Units (mg/dL) Mean Within Within Cone. Day SD* Lab S D " 30.8 0.98 1.12 52.1 0.99 2.14 SI Units (mmol/L) Within Within Mean Day SD* Lab S D " Cone. 0.03 0.03 0.80 1.35 0.03 0.06 Within Lab CV%** 3.7 4.1 No. Observ. 92 91 No. Days 23 23

System VITROS DT60 II

Within Day precision was determined using two runs/day with two replications. Within Lab precision was determined using a single lot of slides and calibrating weekly.

Pub. No. C-354 EN

Version 2.0

INSTRUCTIONS FOR USE


References

HDLC DT
Micro HDL Cholesterol

References
1. 2. 3. 4. 5. Warnick GR, Benderson J, Albers JJ. Dextran Sulfate-Mg+2 Precipitation Procedure for Quantitation of High-Density Lipoprotein Cholesterol. In Cooper GR (ed). Selected Methods of Clinical Chemistry. Washington, D.C.: American Association for Clinical Chemistry; 10:91-99; 1983. NCCLS. Protection of Laboratory Workers from Instrument Biohazards and Infectious Diseases Transmitted by Blood, Body Fluids and Tissue; Approved Guideline. NCCLS Document M29 (OSBN 1-56238). NCCLS, Wayne, PA 19087; 1997. Doumas BT, et al. Differences Between Values for Plasma and Serum in Tests Performed in the Ektachem 700 XR Analyzer, and Evaluation of "Plasma Separator Tubes (PST)." Clin. Chem. 35:151-153; 1989. Calam RR. Specimen Processing Separator Gels: An Update. J Clin Immunoassay. 11:86-90; 1988. Recommendations for Improving Cholesterol Measurement: A Report from the Laboratory Standardization Panel of the National Education Program. US Department of Health and Human Services Public Health Service, National Institutes of Health. NIH Publication No. 90-2964. February 1990. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Venipuncture. NCCLS Document H3. Wayne, PA: NCCLS; 1991. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Skin Puncture. NCCLS Document H4. Wayne, PA: NCCLS; 1991. Clinical Laboratory Handbook for Patient Preparation and Specimen Handling. Fascicle VI: Chemistry/Clinical Microscopy. Northfield, IL: College of American Pathologists; 1992. Warnick GR, Wood PD. National Cholesterol Education Program Recommendations for Measurements of High-Density Lipoprotein Cholesterol: Executive Summary. Clin. Chem. (41)10:1427-1433; 1995. Kimberley MM, Leary ET, Cole TG, Waymack PP. Selection, Validation, Standardization and Performance of a Designated Comparison Method for HDL-Cholesterol for Use in the Cholesterol Reference Laboratory Network. Clin. Chem. (45) 10:1803-1812; 1999. NCCLS. Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition. NCCLS Document C24. Wayne, PA: NCCLS; 1999. Friedewald WT, Levy Rl, Fredrickson DS. Estimation of the Concentration of Low-Density Lipoprotein Cholesterol in Plasma without Use of the Preparative Ultracentrifuge. Clin. Chem. 18:499; 1972. NCEP. Third Report of the National Cholesterol Education Program (NCEP) Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III); Executive Summary. NIH Publication No. 01-3670. National Institutes of Health. Bethesda. Maryland: May, 2001. NCCLS. Interference Testing in Clinical Chemistry. NCCLS Document EP7. Wayne, PA: NCCLS; 1986. Young DS. Effects of Drugs on Clinical Laboratory Tests, ed. 4. Washington D.C.: AACC Press; 1995. Friedman RB, Young DS. Effects of Disease on Clinical Laboratory Tests. Washington, D.C.: AACC Press; 1990. Tryding N, Tufvesson C, Sonntag O (eds). Drug Effects in Clinical Chemistry, ed. 7. Stockholm: The National Corporation of Swedish Pharmacies, Pharmasoft AB, Swedish Society for Clinical Chemistry; 1996. NCCLS. User Evaluation of Precision Performance with Clinical Chemistry Devices. NCCLS Document EP5. Wayne, PA: NCCLS; 1992.

6. 7. 8. 9. 10. 11. 12. 13.

14. 15. 16. 17. 18.

Glossary of Symbols
The Glossary of Symbols table defines the symbols used on the VITROS Chemistry Products packaging and on the VITROS Chemistry Products Assay Sheets. Glossary of Symbols
Do Not Reuse Authorized Representative in the European Community Contains Sufficient for "n" Tests In vitro Diagnostic Medical Device Store At or Below Store At or Above Store Between Consult Instructions for Use Fragile, Handle with Care. Keep Dry

2
SN
REF

Use by or Expiration Date (Year-Month-Day) Lot Number Serial Number Catalog Number or Product Code Attention: See Instructions for Use. Manufacturer

This end up Irritant Manufacturer follows packaging management procedures

A
ml

Version 2.0

Pub. No. C-354 EN

VITRCpS 0

HDLCDT
Micro HDL Cholesterol

INSTRUCTIONS FOR USE


Revision History

Revision History
Date of Revision 2004-02-29 Version 2.0 Description of Technical Changes* > Slide Diagram - changed BaSO2 to BaSO4 > Method Comparison - replaced plots to show all available data > Updated Glossary of Symbols table > New format < > New organization and sections consistent with IVD Directive i> Limitations of the Procedure - updated values for HDL cholesterol concentration and bias in the Known Interferences table > Method Comparison - updated comparison values and plots > Precision - updated all values > References - added all except 1, 8
4

2003-08-11

1.0

The change bars indicate the position of a technical amendment to the text with respect to the previous version of the document.

\AAien this Instructions For Use is replaced, sign and date below and retain as specified by local regulations or laboratory policies, as appropriate.

Signature

Obsolete Date

C
Ortho-Clinical Diagnostics Johnson & Johnson 50-100 Holmers Farm Way High Wycombe Buckinghamshire HP12 4DP United Kingdom Ortho-Clinical Diagnostics, Inc. 100 Indigo Creek Drive Rochester, NY 14626-5101

' Ortho-Clinical Diagnostics


company

VITROS is a trademark of Ortho-Clinical Diagnostics, Inc. Ortho-Clinical Diagnostics, Inc., 2004. All rights reserved.

10

Pub. No. C-354 EN

Version 2.0

VITRCpS
Chemistry I

INSTRUCTIONS FOR USE


VITROS Chemistry Products DT HDL Cholesterol Kit Intended Use

HDLC DT
HDL Cholesterol

For in vitro diagnostic use only. For use in the quantitative measurement of HDL cholesterol (HDLC) concentration in serum and plasma.

Summary and Explanation of the Test


HDL cholesterol is used to evaluate the risk of developing coronary heart disease (CHD). The risk of CHD increases with lower HDL cholesterol concentrations.

Principles of the Procedure


VITROS DT HDL Cholesterol Kit is used to prepare and test samples for high density lipoprotein (HDL) cholesterol on VITROS DT60/DT60 II Chemistry Systems. The specimens must initially be treated with a reagent to remove other lipoproteins which also contain cholesterol, including the very low density (VLDL) and low-density (LDL) classes. The amount of HDL cholesterol can then be determined using VITROS Chemistry Products HDLC DT Slides. HDL is separated by the precipitation of LDL and VLDL using dextran sulfate (MW 50,000) and magnesium chloride 1 that is provided in the VITROS Chemistry Products HDL Tube. The HDL lipoproteins remain in the liquid portion of the tube after centrifugation. This liquid portion is called the supernate and is the portion analyzed. The non-HDL fractions form a pellet on the bottom of the tube and are discarded. A drop of pretreated patient sample is deposited on the slide and is evenly distributed by the spreading layer to the underlying layers. The Triton X-100 (TX100) surfactant in the spreading layer aids in dissociating the cholesterol and cholesterol esters from lipoprotein complexes present in the sample. Hydrolysis of the cholesterol esters to cholesterol is catalyzed by cholesterol ester hydrolase. Free cholesterol is then oxidized in the presence of cholesterol oxidase to form cholestenone and hydrogen peroxide. Finally, hydrogen peroxide oxidizes a leuco dye in the presence of peroxidase to generate a colored dye. The density of dye formed is proportional to the HDL cholesterol concentration present in the pretreated sample and is measured by reflectance spectrophotometry.

Reaction Sequence
high density lipoproteins cholesterol esters + H2O cholesterol + O2 H2O2 + leuco dye Txioo
cholesterol ester hydrolase

cholesterol + cholesterol esters + proteins > cholesterol + fatty acids ^ cholest-4-en-3-one + H2O2 dye + 2H2O

cholesterol oxidase peroxidase

Test Type and Conditions


Test Type and Conditions for HDLC DT
Test Type Colori metric VITROS DT60/DT60 II Module DT60/DT60 II Approximate Incubation Time 5 minutes Temperature 37C (98.6F) Wavelength 660 nm Sample Drop Volume 10 uL

Version 2.0

Pub. No. C-341 EN

HDLC DT
HDL Cholesterol

INSTRUCTIONS FOR USE


Warnings and Precautions

Warnings and Precautions


For in vitro diagnostic use only. Take care when handling materials and samples of human origin. Since no test method can offer complete assurance that infectious agents are absent, consider all clinical specimens, controls, and calibrators potentially infectious. Handle specimens, solid and liquid waste, and test components in accordance with local regulations and NCCLS Guideline M29 2 or other published biohazard safety guidelines. While VITROS HDLC Sample Diluent product is bovine in origin, it should be handled using the same precautions as with any other blood or blood-derived product. For specific warnings and precautions for calibrators, quality control materials, and other components, refer to the Instructions for Use for the appropriate VITROS product, or to other manufacturer's product literature.

Personal Protection and Ventilation


Wear impervious gloves and proper protective clothing. Safety glasses are recommended. Good general ventilation in the work area is recommended. Minimize the potential for production of aerosols.

Accidental Spillage and Disposal


WARNING: VITROS HDL Tubes contain sodium azide. Disposal of reagent into sinks with copper or lead plumbing should be followed with plenty of water to prevent formation of potentially explosive metallic azides.

Absorb spilled material in vermiculite or other suitable absorbents; sweep up and dispose of with clinical waste. Disinfect area with a 5.25% sodium hypochlorite solution (household bleach) diluted with 9 parts water and leave undisturbed for at least 30 minutes. Excess or unwanted materials and packaging for this product should be disposed of with other clinical waste. VITROS HDL Cholesterol Reagent contains sodium azide (0.01%; 0.01 g/dL). Disposal of reagent into sinks with copper or lead plumbing should be followed with copious volumes of water to prevent formation of potentially explosive metallic azides.

First Aid
VITROS HDL Tubes contain gentamicin sulfate and sodium azide. R22 - Harmful if swallowed. Inhalation - Remove to fresh air. Seek medical advice. Skin-Wash skin after each contact with soap and plenty of water for at least 15 minutes. Seek medical attention if skin is cut or punctured. Eye - Immediately flush eyes, including under the eyelids, with plenty of water for at least 15 minutes and get medical attention. Ingestion - Drink 1 -2 glasses of water. Seek medical advice.

Transportation
This product is not classified as a dangerous substance for transportation purposes. However, package with adequate protection to prevent breakage and ship under refrigerated conditions.

Reagents
Slide Diagram

HDLC DT Slide Ingredients


Reactive ingredients per cm
2

-"*

Triton X-100 0.8 mg; cholesterol oxidase {Nocardia, E.C.1.1.3.6) 0.2 U; cholesterol ester hydrolase (Candida rugosa, E.C.3.1.1.13) 0.7 U; peroxidase (horseradish root, E.C.1.11.1.7) 5.3 U; and 2-(3,5-dimethoxy-4-hydroxyphenyl)4,5-bis-(4-dimethylaminophenyl) imidazole (leucodye) 0.2 mg.

Other ingredients
Pigment, binder, buffer, surfactants, stabilizers and cross-linking agent.

1. Jpper slide mount 2. Spreading layer (BaSO4) > Triton X-100 cholesterol ester hydrolase cholesterol oxidase peroxidase leuco dye 3. Sublayer > buffer, pH 6.25 4. Support layer 6. Lower slide mount

HDL Tubes Reactive Ingredients


Dextran sulfate (MW50,000) 0.8 mg, magnesium chloride hexahydrate 6.7 mg

Other ingredients
Sodium azide 0.22%, gentamicin sulfate <0.01%, dye and preservatives

Pub. No. C-341 EN

Version 2.0

INSTRUCTIONS FOR USE


Specimen Requirements

HDLC DT
HDL Cholesterol

HDLC Sample Diluent


Reactive Ingredients
Bovine serum albumin 7%

Other ingredients
Sodium azide 0.05%, inorganic salts and preservatives

Slide Handling
Do not use slides with damaged or incompletely sealed packaging.

Slide Preparation
IMPORTANT: The slide must reach room temperature, 18-28C (64-82F), before the wrapper is opened. Do not use unopened slides that have been at room temperature, 18-28C (64 "-82 F) for >48 hours. 1. Remove the unopened slide from the box. 2. Warm the unopened slide for 15 minutes. Unopened slides that remain in the sealed wrapper may be returned to refrigerator or freezer storage. 3. Remove the packaging and place the slide into the loading station within 15 minutes after opening.

Kit Storage and Stability


VITROS DT HDL Cholesterol Kit components are stable until the expiration date on the carton when they are stored and handled as specified.

Kit Storage and Stability for HDLC DT


Kit Component Slide Refrigerated Frozen Room temperature Room temperature Refrigerated Frozen Sample Diluent* Refrigerated Frozen * Discard if the solution becomes cloudy or turbid. Verify performance with quality control materials: - If the system is turned off for more than 2 hours. 2-8C (36-46F) <-18C (<0F) Until expiration date Until expiration date 2-8C (36-46F) <-18C(<0F) 18-28C(64-82F) 18O-28C(64-82F) 2-8C (36-46F) <-18C(<0F) Until expiration date Until expiration date <48 hours <15 minutes Until expiration date Until expiration date Storage Condition Stability

Opened Tubes

Specimen Requirements
Handle specimens as biohazardous material.

Specimens Recommended
Serum Plasma:3 Heparin Certain collection devices have been reported to affect other analytes and tests." Confirm that your collection devices are compatible with this test. IMPORTANT:

Specimens Not Recommended


. Plasma:5 EDTA

Version 2.0

Pub. No. C-341 EN

HDLCDT
HDL Cholesterol Serum and Plasma Specimen Collection and Preparation
Collect specimens using standard laboratory procedures.6 7

INSTRUCTIONS FOR USE


Specimen Pretreatment and Testing Procedure

Patient Preparation No special patient preparation is necessary unless the HDL cholesterol test is part of a complete lipid profile. Then, a 12- to 14-hour fast is necessary.1 Special Precautions Centrifuge specimens and remove the serum or plasma from the cellular material within 3 hours of collection. If further processing is delayed, store in the refrigerator at 2-8C (36-46F).

Specimen Handling and Storage


- , ' ' ) ( Handle specimens as biohazardous material,

Handle and store specimens in stoppered containers to avoid contamination and evaporation. Mix samples by gentle inversion and bring to room temperature, 18-28C (64-82F), prior to analysis.

Specimen Storage and Stability for HDLC DT: Serum and Plasma '*
Storage Original Specimen Refrigerated Frozen Pretreated Specimen (Supernate) Freezer Temperature 2-8C (36-46F) <-18C(<0F) <-20C (<-4F) <-70C (<-94F) Avoid repeated freeze-thaw cycles. Stability <3 days <4 weeks* <3 months <2 years

For longer storage, separate HDL fractions and freeze the supernate.

IMPORTANT:

Specimen Pretreatment and Testing Procedure


Materials Provided
25 VITROS Chemistry Products HDLC DT Slides 27 VITROS Chemistry Products HDL Tubes VITROS Chemistry Products HDLC Sample Diluent

Materials Required But Not Provided


. VITROS Chemistry Products DT Calibrator Kit VITROS Chemistry Products DT Controls I & II VITROS DT Pipette Centrifuge capable of generating forces of 1500 x g, or a microcentrifuge capable of generating forces of 12,600 x g A vortex mixer is recommended

Special Precautions
Use a vortex speed that will not cause the mixture to spill out of the sample tube.

Procedure
IMPORTANT; Do riot pretreat calibrators.

Be sure to use components from the same kit lot number. IMPORTANT; 1. Allow at least 10 minutes for refrigerated VITROS HDL Tubes and VITROS HDLC Sample Diluent to reach room temperature. If these materials are stored at -18C (0F), they will require a longer warm-up period (at least 30 minutes). Once opened, the VITROS HDLC Sample Diluent should be stored tightly capped in the refrigerator between uses. 2. Pipette serum or heparin plasma to the 0.5 mL mark on the VITROS HDL Tube. 3. Cap and mix thoroughly for 30 seconds - a vortex mixer is recommended. NOTE: 4. The sample will become cloudy during mixing.

Let stand for a minimum of 5 minutes. Pub. No. C-341 EN Version 2.0

INSTRUCTIONS FOR USE


Calibration

HDLC DT
HDL Cholesterol

5. Centrifuge the VITROS HDL Tube for 10 minutes at 1,500 x g. Alternatively, you may centrifuge the tube for a shorter duration using a microcentrifuge for 95 seconds at 12,600 x g. 6. Visually check supemates for clarity. The non-HDL fractions form a pellet on the bottom of the tube. Do not disturb the pellet. If the supernate is clear, transfer cleared supernate directly from the VITROS HDL Tube to a sample cup for analysis on the DT analyzer. IMPORTANT: Supernates should be removed from the pelleted precipitate as soon as possible following centrifugation. The supernates should be used within 15 minutes. If analysis on the VITROS DT60/DT60 II Chemistry System is to be delayed more than one hour, the supernate should be refrigerated. If analysis is delayed longer than 8 hours, refer to the "Specimen Handling and Storage" section.

7. Analyze treated sample as instructed in the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Operating Instructions
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System. Bring all fluids and samples to room temperature, 18-28C (64-82F); analyze immediately. IMPORTANT:

Sample Dilution
If HDL concentrations exceed the system's reportable (dynamic) range or if the supernatant is cloudy or has floating particles after pretreatment: 1. Dilute the untreated sample with an equal volume of VITROS HDLC Sample Diluent. 2. Mix gently by inverting several times. 3. Pipette 0.5 mL of the diluted specimen into a second VITROS HDL Tube. 4. Repeat steps 1 through 6 of the procedure above. Remember to multiply the result by the dilution factor of 2.0. 5. Reanalyze. 6. Multiply the results by 2 to obtain an estimate of the original sample's HDL concentration. MOTE; Additional dilutions are not recommended.

Calibration
Required Calibrators
VITROS Chemistry Products DT Calibrator Kit, bottles 1,2, and 4

Calibrator Preparation, Handling, and Storage


IMPORTANT: Do not pretreat calibrators. Refer to the instructions for use for VITROS DT Calibrator Kit.

Calibration Procedure
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

When to Calibrate
Calibrate: When the slide lot number changes. When critical system parts are replaced due to service or maintenance. When government regulations require. - For example, in the USA, CLIA regulations require calibration or calibration verification at least once every six months. The VITROS HDLC DT slides may also need to be calibrated: If quality control results are consistently outside acceptable range. After certain service procedures have been performed. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Version 2.0

Pub. No. C-341_EN

HDLCDT
HDL Cholesterol

INSTRUCTIONS FOR USE


Quality Control

Calculations
Reflectance from the slide is measured at 660 nm after the fixed incubation time. Once a calibration has been performed for each slide lot, high-density lipoprotein cholesterol concentration in unknown samples can be determined using the softwareresident endpoint colorimetric math model and the response obtained from each unknown test slide.

Validity of a Calibration
Calibration parameters are automatically assessed by the VITROS DT60/DT60 II Chemistry System against a set of quality parameters resident within the analyzer software. Failure to meet any of the pre-defined quality parameters results in a failed calibration. The calibration report should be used in conjunction with quality control results to determine the validity of a calibration.

Reportable (Dynamic) Range Reportable (Dynamic) Range for HDLC DT


Conventional (mg/dL) 1-110 SI Units (mmol/L) 0.03-2.84

For out-of-range samples, refer to "Sample Dilution."

Traceability of the Calibration


Values assigned to the VITROS Chemistry Products DT Calibrator Kit for High Density Lipoprotein-Cholesterol (HDLC) Reagent with VITROS HDLC DT Slides are traceable to the Certified NIST (National Institute of Standards and Technology) Reference Material, SRM (Standard Reference Material) 911b. The Ortho-Clinical Diagnostics Calibration Laboratory uses SRM 911b to assign values to secondary standards for calibration of an HDLC Selected Method. The Selected Method supports VITROS HDL Cholesterol reagent value assignment for the VITROS DT Calibrator Kit. The HDLC Selected Method includes (MW 50,000) Dextran Sulfate precipitation of non-HDL lipoproteins according to the recommendations of the Centers for Disease Control (CDC), 10 followed by automated enzymatic determination of supernate (HDL fraction) cholesterol concentration.

Quality Control
IMPORTANT: Controls must be pretreated.

Procedure Recommendations
Handle quality control materials as biohazardous material. Choose control levels that check the clinically relevant range. Analyze quality control materials in the same manner as patient samples, before or during patient sample processing. To verify system performance, analyze control materials: - After calibration. - According to local regulations or at least once each day that the test is being performed. - After specified service procedures are performed. Refer to the operator's manual for your VITROS DT60/DT60 II System. If control results fall outside your acceptable range, investigate the cause before deciding whether to report patient results. For general quality control recommendations, refer to Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition " or other published guidelines. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Quality Control Material Selection


IMPORTANT; VITROS DT Control I & II are recommended for use with the VITROS DT60/DT60II Chemistry System. Evaluate the performance of other commercial control fluids for compatibility with this test before using for quality control.

Control materials other than VITROS DT Controls I & II may show a difference when compared with other HDL cholesterol methods if they: - Depart from a true human matrix. - Contain high concentrations of preservatives, stabilizers, or other nonphysiological additives. Do not use control materials stabilized with ethylene glycol.

Quality Control Material Preparation and Storage


Refer to the instructions for use for VITROS Chemistry Products DT Control I & II or to other manufacturer's product literature.

Pub. No. C-341 EN

Version 2.0

INSTRUCTIONS FOR USE


Interpretation of Results and Expected Results

HDLC DT
HDL Cholesterol

Interpretation of Results and Expected Results


Interpretation of Results
LDL cholesterol and VLDL cholesterol can be calculated from the results of the VITROS CHOL DT Slide, VITROS TRIG DT Slide, and the results for HDL cholesterol to provide complete lipoprotein profiles. LDL = CHOL-HDLC-VLDL VLDL = TRIG/5 for conventional units (mg/dL) VLDL = TRIG/2.2 for SI units (mmol/L) Calculation of LDL is not appropriate for samples with triglyceride concentrations greater than 400 mg/dL (4.57 mmol/L) or with samples from patients who have type III hyperlipoproteinemia (electrophoresis "broad beta" lipoprotein) present. 12

Expected Results
These guidelines have been recommended by the National Institutes of Health. 13

Reference Interval for HDLC DT


Conv. Units (mg/dL) <40 >60 SI Units (mmol/L) <1.0 >1.6

Low High

Each laboratory should confirm the validity of these intervals for the population it serves.

Reporting Units and Unit Conversion


The VITROS DT60/DT60 II Chemistry System may be programmed to report HDL cholesterol results in conventional and SI units.

Reporting Units and Unit Conversion for HDLC DT Conventional Units SI Units
mg/dL mmol/L (mg/dL x 0.02586)

Limitations of the Procedure


Known Interferences
The VITROS DT HDL Cholesterol Kit method was screened for interfering substances following NCCLS Protocol EP7. " The substances listed in the table, when tested at the concentrations indicated, caused the bias shown.

Known Interfering Substances for HDLC DT


Interferent Interferent* Ascorbic acid Dipyrone Dopamine N-acetylcysteine
* **

HDL Cholesterol Concentration Comments High Therapeutic High IV Drip High IV Drip Oral Therapeutic Conv. (mg/dL) 40.9 40.5 44.6 40.2 SI (mmol/L) 1.06 1.05 1.15 1.04

Bias** Conv. (mg/dL) -7 -7 -9 -6 SI (mmol/L) -0.18 -0.18 -0.22 -0.14

Concentration 170umol/L 3 mg/dL 3.6 mmol/L 12 mg/dL 200 umol/L 4 mg/dL 0.61 mmol/L 10 mg/dL

It is possible that other interfering substances may be encountered. These results are representative; however, your results may differ somewhat due to test-to-test variation. The degree of interference at concentrations other than those listed might not be predictable. The bias is an estimate of the maximum difference observed.

Other Limitations
Certain drugs and clinical conditions are known to alter HDL cholesterol concentration in vivo. For additional information, refer to one of the published summaries.1516'17

Version 2.0

Pub. No. C-341 EN

HDLCDT
HDL Cholesterol

INSTRUCTIONS FOR USE


Performance Characteristics

Performance Characteristics
Method Comparison
The plots and table show the results of a comparison of the VITROS HDL Cholesterol Method analyzed on the VITROS DT60 II Chemistry System using the Dextran Sulfate/Enzymatic 10 comparative method using pretreated serum specimens. The VITROS DT HDL Cholesterol method has met the requirements of the CDC/NCEP Lipid Standardization Program for manufacturers.

Method Comparison for HDLC DT: Serum


Conventional Units y =x - 100 SI Units y =x

3.0 I 2.5
E

1 80 I 60
g 40 20 0 0 20 40 60 80 100
Comparative Method: Dextran Sulfate/Enzymatic (mg/dL)

I 2.0
1.5

o s

I 0.5
5 0.0
0.0 0.5 1.0 1.5 2.0 2.5 3.0
Comparative Method: Dextran Sulfate/Enzymatic (mmol/L)

Method Comparison for HDLC DT: Serum


Conventional Units (mg/dL) n DT60 II System vs. comparative method 43 Correlation Slope Coefficient 1.00 0.998 Range of Sample Cone. Intercept Sy.x 0.02 1.23 SI Units (mmol/L) Range of Sample Cone. Intercept 0.3-2.6 0.00

Sy.x 0.03

12-101

Precision
Precision was evaluated with quality control materials on VITROS the DT60 II System following NCCLS Protocol EP5. 1S The data presented are a representation of test performance and are provided as a guideline. Variables such as sample handling and storage, reagent handling and storage, laboratory environment, and system maintenance can affect reproducibility of test results.

Precision for HDLC DT: Serum


Conventional Units (mg/dL) Mean Within Within Cone. Day SD* Lab SD** 31.5 0.56 1.01 2.14 53.8 0.90 SI Units (mmol/L) Mean Within Within Cone. Day SD* Lab SD** 0.01 0.82 0.03 0.02 1.39 0.06 Within Lab CV%** 3.2 4.0 No. Observ. 99 100 No. Days 25 25

System VITROS DT60 II

Within Day precision was determined using two runs/day with two replications. Within Lab precision was determined using a single lot of slides and calibrating weekly.

Pub. No. C-341_EN

Version 2.0

INSTRUCTIONS FOR USE


References

HDL6 DT
HDL Cholesterol

References
1. 2. 3. 4. 5. Warnick GR, Benderson J, Albers JJ. Dextran Sulfate-Mg+2 Precipitation Procedure for Quantitation of High-Density Lipoprotein Cholesterol. In Cooper GR (ed). Selected Methods of Clinical Chemistry. Washington, D.C.: American Association for Clinical Chemistry, 10:91-99; 1983. NCCLS. Protection of Laboratory Workers from Instrument Biohazards and Infectious Diseases Transmitted by Blood, Body Fluids and Tissue; Approved Guideline. NCCLS Document M29 (OSBN 1-56238). NCCLS, Wayne, PA 19087; 1997. Doumas BT, et al. Differences Between Values for Plasma and Serum in Tests Performed in the Ektachem 700 XR Analyzer, and Evaluation of "Plasma Separator Tubes (PST)." Clin. Chem. 35:151-153; 1989. Calam RR. Specimen Processing Separator Gels: An Update. J Clin Immunoassay. 11:86-90; 1988. Recommendations for Improving Cholesterol Measurement A Report from the Laboratory Standardization Panel of the National Education Program. US Department of Health and Human Services Public Health Service, National Institutes of Health. NIH Publication No. 90-2964. February 1990. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Venipuncture. NCCLS Document H3. Wayne, PA: NCCLS; 1991. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Skin Puncture. NCCLS Document H4. Wayne, PA: NCCLS; 1991. Clinical Laboratory Handbook for Patient Preparation and Specimen Handling. Fascicle VI: Chemistry/Clinical Microscopy. Northfield, IL: College of American Pathologists; 1992. Warnick GR, Wood PD. National Cholesterol Education Program Recommendations for Measurements of High-Density Lipoprotein Cholesterol: Executive Summary. Clin. Chem. (41)10:1427-1433; 1995. Kimberley MM, Leary ET, Cole TG, Waymack PP. Selection, Validation, Standardization and Performance of a Designated Comparison Method for HDL-Cholesterol for Use in the Cholesterol Reference Laboratory Network. Clin. Chem. (45) 10:1803-1812; 1999. NCCLS. Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition. NCCLS Document C24. Wayne, PA: NCCLS; 1999. Friedewald WT, Levy Rl, Fredrickson DS. Estimation of the Concentration of Low-Density Lipoprotein Cholesterol in Plasma without Use of the Preparative Ultracentrifuge. Clin. Chem. 18:499; 1972. NCEP. Third Report of the National Cholesterol Education Program (NCEP) Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III); Executive Summary. NIH Publication No. 01-3670. National Institutes of Health. Bethesda. Maryland: May, 2001. NCCLS. Interference Testing in Clinical Chemistry. NCCLS Document EP7. Wayne, PA: NCCLS; 1986. Young DS. Effects of Drugs on Clinical Laboratory Tests, ed. 4. Washington D.C.: AACC Press; 1995. Friedman RB, Young DS. Effects of Disease on Clinical Laboratory Tests. Washington, D.C.: AACC Press; 1990. Tryding N, Tufvesson C, Sonntag O (eds). Drug Effects in Clinical Chemistry, ed. 7. Stockholm: The National Corporation of Swedish Pharmacies, Pharmasoft AB, Swedish Society for Clinical Chemistry; 1996. NCCLS. User Evaluation of Precision Performance with Clinical Chemistry Devices. NCCLS Document EP5. Wayne, PA: NCCLS; 1992.

6. 7. 8. 9. 10. 11. 12. 13.

14. 15. 16. 17. 18.

Glossary of Symbols
The Glossary of Symbols table defines the symbols used on the VITROS Chemistry Products packaging and on the VITROS Chemistry Products Assay Sheets.

Glossary of Symbols
Do Not Reuse Use by or Expiration Date (Year-Month-Day) Lot Number Authorized Representative in the European Community Contains Sufficient for "n" Tests In vitro Diagnostic Medical Device Store At or Below Store At or Above Store Between Consult Instructions for Use

I t

Fragile, Handle with Care. Keep Dry This end up Irritant Manufacturer follows packaging management procedures

SN
REF

Serial Number Catalog Number or Product Code Attention: See Instructions for Use. Manufacturer

Version 2.0

Pub. No. C-341 EN

VITRCflS 0

HDICDT
HDL Cholesterol

INSTRUCTIONS FOR USE


Revision History

Revision History
Date of Revision 2004-02-29 Version 2.0 Description of Technical Changes* > Slide Diagram - changed BaSO2 to BaSO, Method Comparison - added that method meets requirements of CDC/NCEP Lipid Standardization Program for manufacturers, replaced plots to show all available data > Updated Glossary of Symbols table New format > New organization and sections consistent with IVD Directive > Limitations of the Procedure - updated values for HDL cholesterol concentration and bias in the Known Interferences table Method Comparison - updated comparison values and plots Precision - updated all values > References - added all except 1, 8

2003-08-11

1.0

The change bars indicate the position of a technical amendment to the text with respect to the previous version of the document.

When this Instructions For Use is replaced, sign and date below and retain as specified by local regulations or laboratory policies, as appropriate.

Signature

Obsolete Date

C
EC

Ortho-Clinical Diagnostics Johnson & Johnson 50-100 Holmers Farm Way High Wycombe Buckinghamshire HP12 4DP United Kingdom Ortho-Clinical Diagnostics, Inc. 100 Indigo Creek Drive Rochester, NY 14626-5101

'Ortho-Clinical Diagnostics

VITROS is a trademark of Ortho-Clinical Diagnostics, Inc. Ortho-Clinical Diagnostics, Inc., 2004. All rights reserved.

10

Pub. No. C-341 EN

Version 2.0

I Product*

VITRII3S
Chemistry!

VITROS Chemistry Products K+ DT Slides

INSTRUCTIONS FOR USE

K DT
Potassium

Intended Use
For in vitro diagnostic use only. VITROS K* DT Slides quantitatively measure potassium (K+) concentration in serum and plasma.

Summary and Explanation of the Test


Potassium is the major cation of the intracellular fluid. Measurement of serum potassium is used for evaluation of electrolyte imbalance, cardiac arrhythmias, muscular weakness, hepatic encephalopathy, and renal failure and for the monitoring of ketoacidosis in diabetes mellitus and intravenous fluid replacement therapy. More than 90% of hypertensive patients with aldosteronism have a low K+; a low K* is also common in vomiting, diarrhea, alcoholism, and folic acid deficiency. High K* values occur in rapid K+ infusion, end stage renal failure, hemolysis, trauma, Addison's disease, metabolic acidosis, acute starvation, dehydration, and acute medical emergency.

Principles of the Procedure


The VITROS IC DT Slide method is performed using the VITROS K+ DT Slide and the VITROS Chemistry Products DT Calibrator Kit on VITROS DT60/DT60 II Chemistry Systems. The VITROS K+ DT Slide is a multilayered, analytical element coated on a polyester support that uses direct potentiometry for measurement of ionic potassium. The slide consists of two ion-selective electrodes, each containing valinomycin (an ionophore for potassium), a reference layer, and a silver and a silver chloride layer coated on a polyester support. A drop of patient sample and a drop of VITROS DT Reference Fluid on separate halves of the slide results in migration of both fluids toward the center of the paper bridge. A stable liquid junction is formed connecting the reference electrode to the sample indicator electrode. Each electrode produces an electrical potential in response to the activity of potassium applied to it. The potential difference poised between the two electrodes is proportional to the potassium concentration in the sample.

Test Type and Conditions


Test Type and Conditions for K*
Test Type Potentiometric VITROS DT60/DT60 II Module DTE/DTE II Approximate Incubation Time 90 or 180 seconds* Temperature 25C (77F) Drop Volume Reference Sample: Fluid: 10 uL 10 ML

Assay time is determined by the Calibration Data Module (CDM).

Warnings and Precautions


For in vitro diagnostic use only. Take care when handling materials and samples of human origin. Since no test method can offer complete assurance that infectious agents are absent, consider all clinical specimens, controls, and calibrators potentially infectious. Handle specimens, solid and liquid waste, and test components in accordance with local regulations and NCCLS Guideline M291 or other published biohazard safety guidelines. For specific warnings and precautions for calibrators, quality control materials, and other components, refer to the instructions for use for the appropriate VITROS product, or to other manufacturer's product literature.

Version 1.0

Pub. No. C-306

K DT
Potassium

INSTRUCTIONS FOR USE


Reagents

Reagents
Slide Diagram

Slide Ingredients Reactive ingredients per cm


2

Silver 0.4 mg; silver chloride 0.2 mg; potassium chloride 63 (.ig; and valinomycin 55 \xg.

. . . - - "" *

*3

Other ingredients
Binders, plasticizers, stabilizer, surfactants and nickel.
7

1. Upper frame 2. Paper Bridge 3. Ion-selective membrane valinomycin 4. Reference layer KCI 6. Silver, silver chloride layer 6. Support layer 7. Lower frame

Slide Handling
Do not use slides with damaged or incompletely sealed packaging.

Slide Preparation
fMPQRTANT: The slide must reach room temperature, 18o-28aC(64o-82F), opened. before the wrapper is

Do not use unopened slides that have been at room temperature, 18 -28 C (64 -82F) for >48 hours. 1. 2. Remove the unopened slide from the box. Warm the unopened slide for 15 minutes. Unopened slides that remain in the sealed wrapper may be returned to refrigerator or freezer storage. 3. Remove the packaging and place the slide into the loading station within 15 minutes after opening.

Slide Storage and Stability


VITROS K* DT Slides are stable until the expiration date on the carton when they are stored and handled as specified.

Slide Storage and S t a b i l i t y f o r K D T


Slides Unopened Storage Condition 3 Room temperature 18-28 C (64-82F) Refrigerated 2-8C (36-46F) Frozen <-18C (<0F) Room temperature 18-28 'C (64-82 F) Stability <48 hours Until expiration date Until expiration date <15 minutes

Opened

Specimen Requirements
WARNING: Handle specimens as biohazardous material.

Specimens Recommended
Serum Plasma:2 Heparin Certain collection devices have been reported to affect other analytes and tests.3 Confirm that your collection devices are compatible with this test.

IMPORTANT:

Specimens Not Recommended


EDTA Citrate Fluoride oxalate Do not use hemolyzed specimens. Lysis of only 0.5% of the erythrocytes can increase potassium levels by 0.5 mmol/L.4 Plasma:5

Pub. No. C-306

Version 1.0

INSTRUCTIONS FOR USE


Testing Procedure Serum and Plasma Specimen Collection and Preparation
Collect specimens using standard laboratory procedures.6 7

K DT
Potassium

Patient Preparation The patient should avoid any exercise of the arm or hand before or during collection because opening and closing the fist increases potassium concentration by 10% to 20%.4 Special Precautions Do not draw specimen from an arm receiving an intravenous transfusion. Fibrin clots may cause incomplete sampling of the specimen.8 - Allow specimens to clot completely in order to prevent fibrin clots. - Inspect plasma specimens for fibrin clots. Do not refrigerate the specimen prior to centrifugation because potassium will leak from the red blood cells.6 Centrifuge specimens and remove the serum or plasma from the cellular material within 2 hours of collection.5 Storage

Specimen Handling and " <:

Handle specimens as biohazardous material.

Handle and store specimens in stoppered containers to avoid contamination and evaporation. Mix samples by gentle inversion and bring to room temperature, 18-28C (64-82F), prior to analysis. Specimen Storage and Stability for K* DT: Serum and Plasma 5 Temperature Storage Stability Room temperature 18-28C(64-82F) <6 weeks 2-8C (36-46F) <6 weeks Refrigerated Frozen <-18C (<0F) <1 year

Testing Procedure
Materials Provided
VITROS Chemistry Products K* DT Slides

Materials Required But Not Provided


. VITROS Chemistry Products DT Calibrator Kit Quality control materials, such as VITROS Chemistry Products DT Control I & II VITROS Chemistry Products DT Reference Fluid VITROS DTE Dual Sample Cups VITROS DTE Pipette

Operating Instructions
I I Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System. Bring all fluids and samples to room temperature, 18-28C (64-82"F), prior to analysis. IMPORTANT:

Sample Dilution
Potassium concentrations outside the reportable (dynamic) range are not expected. Diluted samples should not be analyzed with VITROS K* Slides because dilution changes the concentration of solids in plasma water and the ionic strength of the sample.

Version 1.0

Pub. No. C-306

KDT
Potassium

INSTRUCTIONS FOR USE


Calibration

Calibration
Required Calibrators
VITROS Chemistry Products DT Calibrator Kit, bottles 1 and 2

Calibrator Preparation, Handling, and Storage


Refer to the instructions for use for VITROS DT Calibrator Kit.

Calibration Procedure
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System. NOTE: Calibrate potassium in duplicate by running each bottle twice.

When to Calibrate
Calibrate: When the slide lot number changes. When critical system parts are replaced due to service or maintenance. When government regulations require. - For example, in the USA, CLIA regulations require calibration or calibration verification at least once every six months. When the VITROS DT Reference Fluid lot number changes. The VITROS K' DT test may also need to be calibrated: If quality control results are consistently outside acceptable range. After certain service procedures have been performed. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Calculations
The analyzer measures the potential difference in millivolts between the two electrodes of a potentiometric slideone in contact with the sample to be analyzed and the other in contact with the electrolyte reference fluid. A linear relationship exists between the measured potential difference observed on the slide and the logarithm of potassium concentration, i.e. the Nernst equation for ion-selective electrodes. Once the calibration has been established for each slide lot, unknown potassium concentration for a given sample can be determined using the software-resident math model and the measured potential difference.

Validity of a Calibration
Calibration parameters are automatically assessed by the VITROS DT60/DT60II Chemistry System against a set of quality parameters resident within the analyzer software. Failure to meet any of the pre-defined quality parameters results in a failed calibration. The calibration report should be used in conjunction with quality control results to determine the validity of a calibration.

Reportable (Dynamic) Range Reportable (Dynamic) Range for K* DT


Conventional and SI Units (mmol/L)

1.0-11.0

Traceability of the Calibration


Values assigned to the VITROS Chemistry Products DT Calibrator Kit for potassium are traceable to the Certified NIST (National Institute of Standards and Technology) Reference Material, SRM" (Standard Reference Material) 918a. The OrthoClinical Diagnostics calibration laboratory uses SRM 918a to calibrate the flame atomic emission spectroscopy method9 to support potassium value assignment for the VITROS DT Calibrator Kit.

Pub. No. C-306

Version 1.0

INSTRUCTIONS FOR USE


Quality Control

K DT
Potassium

Quality Control
Procedure Recommendations
| Handle quality control materials as biohazardous material. Choose control levels that check the clinically relevant range. Analyze quality control materials in the same manner as patient samples, before or during patient sample processing. To verify system performance, analyze control materials: - After calibration. - According to local regulations or at least once each day that the test is being performed. - After specified service procedures are performed. Refer to the operator's manual for your VITROS DT60/DT60 II System. If control results fall outside your acceptable range, investigate the cause before deciding whether to report patient results. For general quality control recommendations, refer to Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition10 or other published guidelines. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Quality Control Material Selection


I IMPORTANT; VITROS DT Control I & II are recommended for use with the VITROS DT60/DT60 II Chemistry System. Evaluate the performance of other commercial control fluids for compatibility with this test before using for quality control.

Control materials other than VITROS DT Controls I & II may show a difference when compared with other potassium methods if they: - Depart from a true human matrix. - Contain high concentrations of preservatives, stabilizers, or other nonphysiological additives. Do not use control materials stabilized with ethylene glycol.

Quality Control Material Preparation and Storage


Refer to the instructions for use for VITROS Chemistry Products DT Control I & II or to other manufacturer's product literature.

Expected Values and Reporting Units


The serum reference interval is based on an external study.11 The plasma reference interval is based on an external study.4 Reference Interval for K+ Conventional and SI Units Serum Plasma 3.5-5.1 mmol/L 0.1-0.7 mmol/L lower than serum range

Each laboratory should confirm the validity of these intervals for the population it serves.

Reporting Units
The VITROS DT60/DT60 II Chemistry System may be programmed to report potassium results in conventional units.

Reporting Units for K+ DT


Conventional and SI Units mmol/L

Limitations of the Procedure


Known Interferences
None have been identified.

Other Limitations
Certain drugs and clinical conditions are known to alter potassium concentration in vivo. For additional information, refer to one of the published summaries.12 13

Version 1.0

Pub. No. C-306

K DT
Potassium

INSTRUCTIONS FOR USE


Performance Characteristics

Performance Characteristics
Method Comparison

The plot and table show the results of a comparison of the VITROS DT60 II Chemistry System with the VITROS 950 Chemistry

System. Testing followed NCCLS Protocol EP9.14 The table also shows the results of comparisons of the VITROS K* DT slide at an assay time of approximately 90 seconds with the VITROS K* DT slide at an assay time of approximately 180 seconds.

Method Comparison for K+ DT: Serum


Conventional and SI Units 12

5
-

IO 8"

I
to

,.
ft

10

12

Comparative Method: VITROS 950 System (mmol/L)

Method Comparison for K* DT: Serum


Conventional and SI Units (mmol/L) Range of Sample Cone. Intercept Sy.x

n DT60/DT60 II System vs. 950 System 90 seconds vs. 180 seconds 35 35

Slope 1.02 1.02

Correlation Coefficient 0.999 1.000

1.8-7.5 1.6-7.3

-0.20 -0.04

0.06 0.04

Precision
Precision was evaluated with quality control materials on VITROS the DT60 II System following NCCLS Protocol EP5.15 The data presented are a representation of test performance and are provided as a guideline. Variables such as sample handling and storage, reagent handling and storage, laboratory environment, and system maintenance can affect reproducibility of test results.

Precision for K+ DT: Serum


Conventional and SI Units (mmol/L) System VITROS DT60 II Mean Cone. 2.4 5.3 Within Day SD* Within Lab SD** Within Lab CV%** 0.02 0.07 0.04 0.09 1.5 1.7 No. Observ. 88 88 No. Days 22 22

Within Day precision was determined using two runs/day with two replications. Within Lab precision was determined using a single lot of slides and calibrating weekly.

Pub.

No. C-306

Version 1.0

INSTRUCTIONS FOR USE


References

KOT
Potassium

References
1. NCCLS. Protection of Laboratory Workers from Instrument Biohazards and Infectious Diseases Transmitted by Blood, Body Fluids and Tissue; Approved Guideline. NCCLS Document M29 (OSBN 1-56238). NCCLS, Wayne, PA 19087; 1997. Doumas BT, et al. Differences Between Values for Plasma and Serum in Tests Performed in the Ektachem 700 XR Analyzer, and Evaluation of "Plasma Separator Tubes (PST)." Clin. Chem. 35:151-153; 1989. Calam RR. Specimen Processing Separator Gels: An Update. J Clin Immunoassay. 11:86-90; 1988. Tietz NW(ed). Fundamentals of Clinical Chemistry, ed. 3. Philadelphia: WB Saunders; 617; 1987. Clinical Laboratory Handbook for Patient Preparation and Specimen Handling. Fascicle VI: Chemistry/Clinical Microscopy. Northfield, IL: College of American Pathologists; 1992. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Venipuncture. NCCLS Document H3. Wayne, PA: NCCLS; 1991. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Skin Puncture. NCCLS Document H4. Wayne, PA: NCCLS;

1991.
Slockbower JM, Blumenfeld TA (ed.). Collection and Handling of Laboratory Specimens. Philadelphia: Lippincott Co; 201; 1983. 9. Velapoldi R.A., et al. A reference method for the determination of potassium in serum. National Institute of Standards and Technology Special Publication 260-63. Gaithersburg, MD, 1978. 10. NCCLS. Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition. NCCLS Document C24. Wayne, PA: NCCLS; 1999. 11. Tietz NW (ed). Fundamentals of Clinical Chemistry, ed. 4. Philadelphia: WB Saunders; 809; 1996. 12. Young DS. Effects of Drugs on Clinical Laboratory Tests, ed. 4. Washington D.C.: AACC Press; 1995. 13. Friedman RB, Young DS. Effects of Disease on Clinical Laboratory Tests. Washington, D.C.: AACC Press; 1990. 14. NCCLS. Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline. NCCLS Document EP9. Wayne, PA: NCCLS; 1995. 15. NCCLS. User Evaluation of Precision Performance with Clinical Chemistry Devices. NCCLS Document EP5. Wayne, PA: NCCLS;

1992.

Glossary of Symbols
The Glossary of Symbols table defines the symbols used on the VITROS Chemistry Products packaging and on the VITROS Chemistry Products Assay Sheets.

Glossary of Symbols
Do Not Reuse Manufacturer s-e I ma* Authorized Representative Contains Sufficient for "n" Tests For In Vitro Diagnostic Use Store Between Consult Instructions for Use Fragile, Handle with Care. Keep Dry This end up Use by or Expiration Date (YYYY-MM-DD) Lot Number Manufacturer's Serial Number Catalog Number or Product Code Attention: See Instructions for Use.

X 1

Store At or Below Store At or Above

Version 1.0

Pub. No. C-306

KDT
Potassium

INSTRUCTIONS FOR USE


Revision History

Revision History
Date of 'Revision 2003-03-28 Description of Technical Changes* New format New organization and sections consistent with IVD Directive > Specimens Not Recommended - added citrate and fluoride oxalate > Patient Preparation - updated > Special Precautions - updated Specimen Storage and Stability - updated stability values Quality Control Material Selection - added the statement regarding ethylene glycol Expected Values and Reporting Units - updated values Reference Interval - updated values Method Comparison - updated all data and the plot Precision - updated all data > References - added all The change bars indicate the position of a technical amendment to the text with respect to the previous version of the document. Version 1.0

When this Instructions For Use is replaced, sign and date below and retain as specified by local regulations or laboratory policies, as appropriate.

Signature

Obsolete Date

C
EC

Ortho-Clinical Diagnostics Mandeville House 62 The Broadway Amersham Buckinghamshire HP7 OHJ England Ortho-Clinical Diagnostics, Inc. 100 Indigo Creek Drive Rochester, NY 14626-5101

' Otto-Clinical Diagnostics


aH company

VITROS is a trademark of Ortho-Clinical Diagnostics, Inc. Ortho-Clinical Diagnostics, Inc., 2003. All rights reserved. Printed in USA.

Pub. No. C-306

Version 1.0

Chcmbtry

INSTRUCTIONS FOR USE


VITROS Chemistry Products LAC DT Slides Intended Use
For in vitro diagnostic use only. VITROS LAC DT Slides quantitatively measure lactate (LAC) concentration in plasma.

1ACDT
Lactate

Summary and Explanation of the Test


Lactate is the end product of the anaerobic metabolism of glucose. The concentration of lactate in the blood is dependent on the rate of production in muscle cells and erythrocytes and the rate of metabolism in the liver. Lactic acidosis usually results from overproduction or underutilization of lactate. Elevated lactate levels can occur as a result of tissue hypoxia; diabetes mellitus; phenformin therapy; malignancies; glycogen storage disease; ethanol, methanol, or salicylate ingestion; and metabolic acidosis.1

Principles of the Procedure


The VITROS LAC DT Slide method is performed using the VITROS LAC DT Slide and the VITROS Chemistry Products DT Calibrator Kit on VITROS DT60/DT60 II Chemistry Systems. The VITROS LAC DT Slide is a multilayered, analytical element coated on a polyester support. A drop of patient sample is deposited on the slide and is evenly distributed by the spreading layer to the underlying layers. Lactate in the sample is oxidized by lactate oxidase to pyruvate and hydrogen peroxide. The hydrogen peroxide generated oxidizes the 4-aminoantipyrine, 1,7-dihydroxynaphthalene dye system in a horseradish-peroxidase-catalyzed reaction and results in a dye complex. The slide is incubated and the intensity of the dye complex is measured spectrophotometrically.

Reaction Sequence
L-(+)-lactic acid + O2
lactate oxidase

->

pyruvate + H2O2
peroxidase

2H2O2 + 4-aminoantipyrine + 1,7-dihydroxynaphthalene

red dye

Test Type and Conditions


Test Type and Conditions for LAC DT
Test Type Colorimetric VITROS DT60/DT60 II Module DT60/DT60 II Approximate Incubation Time 5 minutes Temperature 37C (98.6F) Wavelength 555 nm Sample Drop Volume 10 uL

Warnings and Precautions


For in vitro diagnostic use only. Take care when handling materials and samples of human origin. Since no test method can offer complete assurance that infectious agents are absent, consider all clinical specimens, controls, and calibrators potentially infectious. Handle specimens, solid and liquid waste, and test components in accordance with local regulations and NCCLS Guideline M292 or other published biohazard safety guidelines. For specific warnings and precautions for calibrators, quality control materials, and other components, refer to the instructions for use for the appropriate VITROS product, or to other manufacturer's product literature.

Version 1.0

Pub. No. C-357

LACDT
Lactate

INSTRUCTIONS FOR USE


Reagents

Reagents
Slide Diagram

Slide Ingredients

Reactive ingredients per cm

, 2

Lactate oxidase (Pediococcus sp., E.C.1.1.3.2) 0.3 U; peroxidase (horseradish


" ' '' - - *

root, E.C.1.11.1.7) 1.5 U; 1,7-dihydroxynapthalene (dye precursor) 43 ug; and 4-aminoantipyrine hydrochloride (dye precursor) 65 ug.

~*~~
. -

4
S

1. Upper slide mount 2. Spreading layer (TIO2) 3. Reagent layer lactate oxidase peroxidase dye precursors buffer, pH 6.25 4. Support layer 6. Lower slide mount

Other ingredients
Pigment, binders, buffer, stabilizers, cross-linking agent, surfactants and enzyme cofactor.

Slide Handling
Do not use slides with damaged or incompletely sealed packaging.

Slide Preparation
The slide must reach room temperature, 18-28C (64 -82 F), before the wrapper is opened. Do not use unopened slides that have been at room temperature, 78-28<C (64 -82 F) for >48 hours. 1. 2. Remove the unopened slide from the box. Warm the unopened slide for 15 minutes. Unopened slides that remain in the sealed wrapper may be returned to refrigerator or freezer storage. 3. Remove the packaging and place the slide into the loading station within 15 minutes after opening.

Slide Storage and Stability


VITROS LAC DT Slides are stable until the expiration date on the carton when they are stored and handled as specified.

Slide Storage and Stability for LAC DT


Slides Unopened Opened Storage Condition Room temperature 18-28C (64-82F) Frozen <-18C (<0F) Room temperature 18-28C (64-82F) Stability <48 hours Until expiration date <15 minutes

Specimen Requirements
WARN I M(k Handle specimens as biohazardous material.

Specimens Recommended
Plasma:3 Fluoride oxalate Heparinized plasma is acceptable, but precautions must be taken to retard glycoiysis by keeping the whole blood on ice." Certain collection devices have been reported to affect other analytes and tests.5 Confirm that your collection devices are compatible with this test.

IMPORTANT:

Plasma
Specimen Collection and Preparation
Collect specimens using standard laboratory procedures.6 7 Patient Preparation Venous specimens should be obtained without the use of a tourniquet or immediately after the tourniquet is applied. Alternatively, the tourniquet should be removed after the puncture has been performed, and the blood should be allowed to circulate for several minutes before the sample is withdrawn.4 The patient should avoid any exercise of the arm or hand before or during collection of the specimen.4

Pub. No. C-357

Version 1.0

INSTRUCTIONS FOR USE


Testing Procedure

LAC DT
Lactate

Special Precautions For fluoride oxalate plasma, specimens must be collected in tubes that are at least half full. Smaller volume's can result in negative biases. Centrifuge specimens and remove the plasma from the cellular material within 15 minutes of collection.3 Specimen Handling and Storage

Handle specimens as biohazardous material. Handle and store specimens in stoppered containers to avoid contamination and evaporation. Mix samples by gentle inversion and bring to room temperature, 18-28C (64-82F), prior to analysis.

Specimen Storage and Stability for LAC DT: Plasma3 Storage Temperature
Room temperature Refrigerated Frozen 18O-28C(64O-82F) 2-8C (36-46F) <-18C(<0F)

Stability <8 hours <14days <1 month

Testing Procedure
Materials Provided
VITROS Chemistry Products LAC DT Slides

Materials Required But Not Provided


. VITROS Chemistry Products DT Calibrator Kit Quality control materials, such as VITROS Chemistry Products DT Control I & II Isotonic saline or reagent-grade water VITROS DT Pipette

Operating Instructions
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System. Bring all fluids and samples to room temperature, 18-28C (64-82'F), prior to analysis. IMPORTANT:

Sample Dilution
If lactate concentrations exceed the system's reportable (dynamic) range: 1. Dilute the sample with isotonic saline or reagent-grade water. 2. Reanalyze. 3. Multiply the results by the dilution factor to obtain an estimate of the original sample's lactate concentration.

Calibration
Required Calibrators
VITROS Chemistry Products DT Calibrator Kit, bottles 1, 2, and 3

Calibrator Preparation, Handling, and Storage


Refer to the instructions for use for VITROS DT Calibrator Kit.

Calibration Procedure
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

When to Calibrate
Calibrate: When the slide lot number changes. When critical system parts are replaced due to service or maintenance. When government regulations require. - For example, in the USA, CLIA regulations require calibration or calibration verification at least once every six months.

Version 1.0

Pub. No. C-357

LACDT
Lactate

INSTRUCTIONS FOR USE


Quality Control
The VITROS LAC DT test may also need to be calibrated: If quality control results are consistently outside acceptable range. After certain service procedures have been performed. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Calculations
Reflectance from the slide is measured at 555 nm after the fixed incubation time. Once a calibration has been performed for each slide lot, lactate concentration in unknown samples can be determined using the software-resident endpoint colorimetric math model and the response obtained from each unknown test slide.

Validity of a Calibration
Calibration parameters are automatically assessed by the VITROS DT60/DT60 II Chemistry System against a set of quality parameters resident within the analyzer software. Failure to meet any of the pre-defined quality parameters results in a failed calibration. The calibration report should be used in conjunction with quality control results to determine the validity of a calibration.

Reportable (Dynamic) Range Reportable (Dynamic) Range for LAC DT


Conventional and SI Units (mmol/L)

0.5-12.0
For out-of-range samples, refer to "Sample Dilution."

Traceability of the Calibration


I | Values assigned to the VITROS Chemistry Products DT Calibrator Kit for lactate are traceable to gravimetrically prepared standards made from reagent grade lactic acid. The Ortho-Clinical Diagnostics calibration laboratory uses the gravimetrically prepared standard to calibrate an HPLC method for lactate8 to support value assignment for the VITROS DT Calibrator Kit.

Quality Control
Procedure Recommendations
| V ' A R ui Nc: Handle quality control materials as biohazardous material. Choose control levels that check the clinically relevant range. Analyze quality control materials in the same manner as patient samples, before or during patient sample processing. To verify system performance, analyze control materials: - After calibration. - According to local regulations or at least once each day that the test is being performed. - After specified service procedures are performed. Refer to the operator's manual for your VITROS DT60/DT60II System. If control results fall outside your acceptable range, investigate the cause before deciding whether to report patient results. For general quality control recommendations, refer to Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition3 or other published guidelines. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Quality Control Material Selection


I IMPORTANT; VITROS DT Control I & II are recommended for use with the VITROS DT60/DT60 II Chemistry System. Evaluate the performance of other commercial control fluids for compatibility with this test before using for quality control.

Control materials other than VITROS DT Controls I & II may show a difference when compared with other lactate methods if they: - Depart from a true human matrix. - Contain high concentrations of preservatives, stabilizers, or other nonphysiological additives. Do not use control materials stabilized with ethylene glycol.

Quality Control Material Preparation and Storage


Refer to the instructions for use for VITROS Chemistry Products DT Control I & II or to other manufacturer's product literature.

Pub.

No. C-357

Version 1.0

INSTRUCTIONS FOR USE


Expected Values and Reporting Units

UCDT
Lactate

Expected Values and Reporting Units


Reference Interval
This reference interval is the central 95% of results from a study of 168 apparently healthy adults from a working population. The values for heparinized plasma may be higher because lactate rapidly increases in blood as a result of glycolysis.10 Reference Interval for LAC DT Conventional and SI Units (mmol/L)

0.7-2.1
Each laboratory should confirm the validity of these intervals for the population it serves.

Reporting Units
The VITROS DT60/DT60 II Chemistry System may be programmed to report lactate results in conventional and SI units. Reporting Units for LAC DT Conventional and SI Units mmol/L

Limitations of the Procedure


Known Interferences
None identified.

Other Limitations
Certain drugs and clinical conditions are known to alter lactate concentration in vivo. For additional information, refer to one of the published summaries.1112

Performance Characteristics
Method Comparison
I I The plot and table show the results of a comparison of samples analyzed on the VITROS DT60 II System with those analyzed using the VITROS 950 System. Method Comparison for LAC DT: Plasma Conventional and SI Units 12

I
E

Hi8-j
6

I
=
&

,j

o o
a:

4-

10

Comparative Method: VITROS 950 System (mmol/L)

Method Comparison for LAC DT: Plasma Conventional and SIUnits (mmol/L) n DT60 II System vs. 950 System 58 Slope 1.00 Correlation Coefficient 1.000 Range of Sample Activity 0.8-10.3 Intercept 0.01
Sy.x

0.08

Version 1.0

Pub.

No. C-357

1ACDT
Lactate Precision

INSTRUCTIONS FOR USE


References
Precision was evaluated with quality control materials on the VITROS DT60 II System following NCCLS Protocol EP5.13 The data presented are a representation of test performance and are provided as a guideline. Variables such as sample handling and storage, reagent handling and storage, laboratory environment, and system maintenance can affect reproducibility of test results.

Precision for LAC DT: Plasma


Conventional and SI Units (mmol/L) System VITROS DT60 II Mean Cone. 1.6 4.0 Within Day SD* 0.02 0.04 Within Lab SD** 0.03 0.06 Within Lab CV%** 1.6 1.6 No. Observ. 84 83 No. Days 21 21

Within Day precision was determined using two runs/day with two replications. Within Lab precision was determined using a single lot of slides and calibrating weekly.

References
Oliva PB. Am. J. Med. 48:209-225; 1970. NCCLS. Protection of Laboratory Workers from Instrument Biohazards and Infectious Diseases Transmitted by Blood, Body Fluids and Tissue; Approved Guideline. NCCLS Document M29 (OSBN 1-56238). NCCLS, Wayne, PA 19087; 1997. 3. Clinical Laboratory Handbook for Patient Preparation and Specimen Handling. Fascicle VI: Chemistry/Clinical Microscopy. Northfield, IL: College of American Pathologists; 1992. 4. Tietz NW (ed). Fundamentals of Clinical Chemistry, ed. 5. Philadelphia: WB Saunders; 451; 2001. 5. Calam RR. Specimen Processing Separator Gels: An Update. J Clin Immunoassay. 11:86-90; 1988. 6. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Venipuncture. NCCLS Document H3. Wayne, PA: NCCLS; 1991. 7. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Skin Puncture. NCCLS Document H4. Wayne, PA: NCCLS; 1991. 8. Smith JW, Ambrose RT. Determination of Lactic Acid in Human Serum by Ion Exchange Chromatography. Internal Eastman Kodak Company Report. 1982. 9. NCCLS. Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition. NCCLS Document C24. Wayne, PA: NCCLS; 1999. 10. Tietz NW (ed). Fundamentals of Clinical Chemistry, ed. 2. Philadelphia: WB Saunders; 940; 1976. 11. Young DS. Effects of Drugs on Clinical Laboratory Tests, ed. 4. Washington D.C.: AACC Press; 1995. 12. Friedman RB, Young DS. Effects of Disease on Clinical Laboratory Tests. Washington, D C : AACC Press; 1990. 13. NCCLS. User Evaluation of Precision Performance with Clinical Chemistry Devices. NCCLS Document EP5. Wayne, PA: NCCLS; 1992. 1. 2.

Pub. No. C-357

Version 1.0

gj VITFIJ

INSTRUCTIONS FOR USE


Glossary of Symbols

IACDT
Lactate

Glossary of Symbols
The Glossary of Symbols table defines the symbols used on the VITROS Chemistry Products packaging and on the VITROS Chemistry Products Assay Sheets.

Glossary of Symbols
Do Not Reuse Use by or Expiration Date (YYYY-MM-DD) Lot Number ee I KEI> I Manufacturer Authorized Representative Contains Sufficient for "n" Tests For In Vitro Diagnostic Use Store At or Below X Store At or Above Store Between Consult Instructions for Use

I
M

Fragile, Handle with Care. Keep Dry This end up

SN
REF|

Manufacturer's Serial Number Catalog Number or Product Code Attention: See Instructions for Use.

Revision History
Date of Revision 2003-10-01 Version 1.0 Description of Technical Changes* > New format New organization and sections consistent with IVD Directive Test Type and Conditions - updated the wavelength > Reference Interval - updated text > Method comparison - updated all data and the plot > Precision - updated all values > References - added all except 1,10

The change bars indicate the position of a technical amendment to the text with respect to the previous version of the document.

When this Instructions For Use is replaced, sign and date below and retain as specified by local regulations or laboratory policies, as appropriate.

Signature

Obsolete Date

Version 1.0

Pub. No. C-357

LACDT
Lactate

INSTRUCTIONS FOR USE


Revision History

C
EC I REP

Ortho-Clinical Diagnostics Mandeville House 62 The Broadway Amersham Buckinghamshire HP7 OHJ England Ortho-Clinical Diagnostics, Inc. 100 Indigo Creek Drive Rochester, NY 14626-5101

'Ortho-Clinical Diagnostics
company

ill

VITROS is a trademark of Ortho-Clinical Diagnostics, Inc. Ortho-Clinical Diagnostics, Inc., 2003. All rights reserved. Printed in USA.

Pub. No. C-357

Version 1.0

[ Prod wets;

ViTRCpS
Cbemittryl

INSTRUCTIONS FOR USE


VITROS Chemistry Products LDH DT Slides Intended Use

LDHDT
Lactate Dehydrogenase

For in vitro diagnostic use only. VITROS LDH DT Slides quantitatively measure lactate dehydrogenase (LDH) activity in serum and plasma.

Summary and Explanation of the Test


Lactate dehydrogenase is an enzyme present in the cytosol of all human cells; it catalyzes the reversible reduction of pyruvate to lactate using NADH. Causes of high LDH include neoplastic states, hypoxic cardiorespiratory diseases, myocardial infarctions, hemolytic anemias, megaloblastic anemias, hepatic cirrhosis, renal infarction, trauma, muscle damage, muscular dystrophy, shock, and hypotension. In myocardial infarction cases, LDH begins to rise within about 12 hours after infarction and usually returns to normal after two to five days.1

Principles of the Procedure


The VITROS LDH DT Slide method is performed using the VITROS LDH DT Slide and the VITROS Chemistry Products DT Calibrator Kit on VITROS DT60/DT60 II Chemistry Systems. The VITROS LDH DT Slide is a multilayered, analytical element coated on a polyester support. A drop of patient sample is deposited on the slide and is evenly distributed by the spreading layer to the underlying layers. Lactate dehydrogenase catalyzes the conversion of pyruvate and NADH to lactate and NAD+. The oxidation of NADH, which is monitored by reflectance spectrophotometry, is used to measure lactate dehydrogenase activity.

Reaction Sequence
pyruvate + NADH + H+
LDH

- > lactate + NAD+

Test Type and Conditions


Test Type and Conditions for LDH DT
Test Type Rate VITROS DT60/DT60 II Module DTSC/DTSC II Approximate Incubation Time 5 minutes Temperature 37C (98.6F) Wavelength 340 nm Sample Drop Volume 10 uL

Warnings and Precautions


For in vitro diagnostic use only. Take care when handling materials and samples of human origin. Since no test method can offer complete assurance that infectious agents are absent, consider all clinical specimens, controls, and calibrators potentially infectious. Handle specimens, solid and liquid waste, and test components in accordance with local regulations and NCCLS Guideline M292 or other published biohazard safety guidelines. For specific warnings and precautions for calibrators, quality control materials, and other components, refer to the instructions for use for the appropriate VITROS product, or to other manufacturer's product literature.

Version 1.0

Pub. No. C-344

LDHDT
Lactate Dehydrogenase

INSTRUCTIONS FOR USE


Reagents

Reagents
Slide Diagram

Slide Ingredients
Reactive ingredients per cm Other ingredients
Polymer beads, binders, buffer, surfactants, cross-linking agent and stabilizer.
2

Nicotinamide adenine dinucleotide, reduced 44 ug and sodium pyruvate 17 ug.

1. Upper slide mount 2. Spreading layer (beads) sodium pyruvate 3. Reagent layer buffer, pH 7.25 NADH 4. Support layer 5. Lower slide mount

Slide Handling
CAUTION: Do not use slides with damaged or incompletely sealed packaging.

Slide Preparation
IMPORTANT: The slide must reach room temperature, 78-28 C (64-82F), before the wrapper is opened. Do not use unopened slides that have been at room temperature, 18-28 C (64-82F) for >48 hours. 1. 2. Remove the unopened slide from the box. Warm the unopened slide for 15 minutes. Unopened slides that remain in the sealed wrapper may be returned to refrigerator or freezer storage. 3. Remove the packaging and place the slide into the loading station within 15 minutes after opening.

Slide Storage and Stability


VITROS LDH DT Slides are stable until the expiration date on the carton when they are stored and handled as specified.

Slide Storage and Stability for LDH DT


Slides Unopened

Opened

Storage Condition Room temperature 18-28C (64-82F) Refrigerated 2-8C (36-46F) Frozen <-18C (<0F) Room temperature 18-28C (64-82F)

Stability <48 hours Until expiration date Until expiration date <15 minutes

Specimen Requirements
WARMS: Handle specimens as biohazardous material.

Specimens Recommended
I Serum Plasma:3 Heparin Serum and heparin plasma specimens produce similar LDH results on VITROS Systems. Some other methods, however, have shown substantial differences between serum and plasma results due to contamination by platelets in plasma separated by low-speed centrlfugation.'1 The VITROS LDH DT Slide is insensitive to LDH contained within intact platelets;5 therefore, LDH results in comparative methods may not agree with the VITROS System results for heparin plasma specimens. Certain collection devices have been reported to affect other analytes and tests.6 Confirm that your collection devices are compatible with this test.

NOTE:

Specimens Not Recommended


Do not use hemolyzed specimens.17

Pub. No. C-344

Version 1.0

|Sj VITRj

INSTRUCTIONS FOR USE


Testing Procedure Serum and Plasma Specimen Collection and Preparation
Collect specimens using standard laboratory procedures.8'9 Patient Preparation No special patient preparation is necessary. Special Precautions Centrifuge specimens and remove the serum from the cellular material within 1 hour of collection.10

IDH DT
Lactate Dehydrogenase

Specimen Handling and Storage G: Handle specimens as biohazardous material.

Handle and store specimens in stoppered containers to avoid contamination and evaporation. Mix samples by gentle inversion and bring to room temperature, 18-28C (64-82F), prior to analysis.

Specimen Storage and Stability for LDH DT: Serum and Plasma1
Storage Room temperature Refrigerated Frozen Temperature 18O-28C(64-82F) 2-8C (36-46F) <-18C(<0F) Stability <2 days Not recommended* Not recommended*

* LD4 and LD5 isoenzymes are labile at refrigerator and freezer temperatures.

Testing Procedure
Materials Provided
VITROS Chemistry Products LDH DT Slides

Materials Required But Not Provided


. VITROS Chemistry Products DT Calibrator Kit Quality control materials, such as VITROS Chemistry Products DT Control I & II VITROS Chemistry Products 7% BSA or isotonic saline VITROS DT Pipette

Operating Instructions
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System. Bring all fluids and samples to room temperature, 18-28C (64-82F), prior to analysis.

I
|

MPORTANT;

Sample Dilution
If lactate dehydrogenase activities exceed the system's reportable (dynamic) range: 1. Dilute the sample with VITROS 7% BSA or isotonic saline. 2. Reanalyze. 3. Multiply the results by the dilution factor to obtain an estimate of the original sample's lactate dehydrogenase activity.

Calibration
Required Calibrators
VITROS Chemistry Products DT Calibrator Kit, bottles 1, 2, and 4

Calibrator Preparation, Handling, and Storage


Refer to the instructions for use for VITROS DT Calibrator Kit.

Calibration Procedure
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Version 1.0

Pub. No. C-344

1DHDT
Lactate Dehydrogenase

INSTRUCTIONS FOR USE


Quality Control
Calibrate: When the slide lot number changes. When critical system parts are replaced due to service or maintenance. When government regulations require. - For example, in the USA, CLIA regulations require calibration or calibration verification at least once every six months.

When to Calibrate

The VITROS LDH DT test may also need to be calibrated: If quality control results are consistently outside acceptable range. After certain service procedures have been performed. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Calculations
Based on sequential readings of the slide's reflectance at 340 nm over the defined incubation period, a rate of change in reflectance is determined. This rate is used in the software-resident multi-point rate calibration model to compute enzyme activity. Once a calibration has been performed for each slide lot, lactate dehydrogenase activity in unknown samples can be determined from the rate of change in reflectance measured for each unknown test slide.

Validity of a Calibration
Calibration parameters are automatically assessed by the VITROS DT60/DT60 II Chemistry System against a set of quality parameters resident within the analyzer software. Failure to meet any of the pre-defined quality parameters results in a failed calibration. The calibration report should be used in conjunction with quality control results to determine the validity of a calibration.

Reportable (Dynamic) Range Reportable (Dynamic) Range for LDH DT


Conventional and SI Units (U/L) 100-1750 For out-of-range samples, refer to "Sample Dilution."

Traceability of the Calibration

I
|

Values assigned to the VITROS Chemistry Products DT Calibrator Kit for lactate dehydrogenase are traceable to the pyruvateto-lactate (P->L) (Buhl) total lactate dehydrogenase method,12 adapted to a centrifugal analyzer at 37C.

Quality Control
Procedure Recommendations
Handle quality control materials as biohazardous material. Choose control levels that check the clinically relevant range. Analyze quality control materials in the same manner as patient samples, before or during patient sample processing. To verify system performance, analyze control materials: - After calibration. - According to local regulations or at least once each day that the test is being performed. - After specified service procedures are performed. Refer to the operator's manual for your VITROS DT60/DT60 II System. If control results fall outside your acceptable range, investigate the cause before deciding whether to report patient results. For general quality control recommendations, refer to Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition13 or other published guidelines. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Pub. No. C-344

Version 1.0

INSTRUCTIONS FOR USE


Expected Values and Reporting Units

LDH DT
Lactate Dehydrogenase

Quality Control Material Selection

I
| I

IMPORTANT;

VITROS DT Control I & II are recommended for use with the VITROS DT60/DT60 II

Chemistry System. Evaluate the performance of other commercial control fluids for compatibility with this test before using for quality control. Control materials other than VITROS DT Controls I & II may show a difference when compared with other lactate dehydrogenase methods if they: - Depart from a true human matrix. - Contain high concentrations of preservatives, stabilizers, or other nonphysiological additives. Enzyme activity might also vary with enzyme source, diluent temperature, and activation time during reconstitution. Do not use control materials stabilized with ethylene glycol.

Quality Control Material Preparation and Storage


Refer to the instructions for use for VITROS Chemistry Products DT Control I & II or to other manufacturer's product literature.

Expected Values and Reporting Units


Reference Interval
These reference intervals are the central 95% of results from an internal study of 557 apparently healthy individuals. No significant differences between results from the male and female populations were observed.

Reference Interval for LDH DT


Conventional and SI Units (U/L) 313-618 Each laboratory should confirm the validity of these intervals for the population it serves.

Reporting Units
The VITROS DT60/DT60 II Chemistry System may be programmed to report lactate dehydrogenase results in conventional and SI units.

Reporting Units for LDH DT


Conventional and SI Units U/L

Limitations of the Procedure


Known Interferences
None identified.

Other Limitations
Certain drugs and clinical conditions are known to alter lactate dehydrogenase activity in vivo. For additional information, refer to one of the published summaries.1415

Version 1.0

Pub. No. C-344

LDHDT
Lactate Dehydrogenase

INSTRUCTIONS FOR USE


Performance Characteristics

Performance Characteristics
Method Comparison

The plot and table show the results of a comparison of samples analyzed on the VITROS DT60 II System with those analyzed using the VITROS 950 System. Method Comparison for LDH OT: Serum Conventional and SI Units 2000 1600 1200 800 400

400

800

1200

1600

2000

Comparative Method: VITROS 950 System (U/L)

Method Comparison for LDH DT: Serum Conventional and SIUnits (U/L) n DT60 II System vs. 9S0 System 61 Slope 1.03 Correlation Coefficient 0.999 Range of Sample Activity 122-1418 Intercept -13.36 Sy.x 14.94

Precision
Precision was evaluated with quality control materials on the VITROS DT60 II System following NCCLS Protocol EP5.16 The data presented are a representation of test performance and are provided as a guideline. Variables such as sample handling and storage, reagent handling and storage, laboratory environment, and system maintenance can affect reproducibility of test results. Precision for LDH DT: Serum Conventional and SI Units (U/L) System VITROS DT60 II Mean Activity 458 1369 Within Day SD* 6.2 20.9 Within Lab S D " Within Lab CV%** No. Observ. 7.5 23.8 1.6 1.7 84 84 No. Days 21 21

Within Day precision was determined using two runs/day with two replications. Within Lab precision was determined using a single lot of slides and calibrating weekly.

Pub. No. C-344

Version 1.0

INSTRUCTIONS FOR USE


References

LDHDT
Lactate Dehydrogenase

References
1. 2. 3. 4. 5. 6. 7. 8. 9. 10. Tietz NW(ed). Fundamentals of Clinical Chemistry, ed. 3. Philadelphia: WB Saunders; 380-384; 1987. NCCLS. Protection of Laboratory Workers from Instrument Biohazards and Infectious Diseases Transmitted by Blood, Body Fluids and Tissue; Approved Guideline. NCCLS Document M29 (OSBN 1-56238). NCCLS, Wayne, PA 19087; 1997. Doumas BT, et al. Differences Between Values for Plasma and Serum in Tests Performed in the Ektachem 700 XR Analyzer, and Evaluation of "Plasma Separator Tubes (PST)." Clin. Chem. 35:151-153; 1989. Peake MJ, et al. Mechanism of Platelet Interference with Measurement of Lactate Dehydrogenase Activity in Plasma. Clin. .Chem. 30:518-520; 1984. Greenberg N, Byrne D. Plasma Lactate Dehydrogenase Activity Assayed with the Kodak Ektachem 700 Analyzer Is Unaffected by Platelet Contamination. Clin. Chem. 31:1022; 1985. Calam RR. Specimen Processing Separator Gels: An Update. J Clin Immunoassay. 11:86-90; 1988. Young DS. Effects of Preanalytical Variables on Clinical Laboratory Tests. Ed. 2. Washington D C : AACC Press; 3-335, 1997. NCCLS. Procedures forthe Collection of Diagnostic Blood Specimens by Venipuncture. NCCLS Document H3. Wayne, PA: NCCLS;

1991.
NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Skin Puncture. NCCLS Document H4. Wayne, PA: NCCLS;

1991.
Clinical Laboratory Handbook for Patient Preparation and Specimen Handling. Fascicle VI: Chemistry/Clinical Microscopy. Northfield, IL: College of American Pathologists; 1992. 11. Tietz NW. Tietz Textbook of Clinical Chemistry, ed. 3. Philadelphia: WB Saunders; 671; 1999. 12. Buhl SN, Jackson KY, Graffunder B. Optimal Reaction Conditions for Assaying Human Lactate Dehydrogenase Pyruvate-to-Lactate at 20, 30, and 37C. Clin. Chem. 24:261-266; 1978. 13. NCCLS. Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition. NCCLS Document C24. Wayne, PA: NCCLS; 1999. 14. Young DS. Effects of Drugs on Clinical Laboratory Tests, ed. 4. Washington D.C.: AACC Press; 1995. 15. Friedman RB, Young DS. Effects of Disease on Clinical Laboratory Tests. Washington, D.C.: AACC Press; 1990. 16. NCCLS. User Evaluation of Precision Performance with Clinical Chemistry Devices. NCCLS Document EP5. Wayne, PA: NCCLS;

1992.

Glossary of Symbols
The Glossary of Symbols table defines the symbols used on the VITROS Chemistry Products packaging and on the VITROS Chemistry Products Assay Sheets.

Glossary of Symbols
Do Not Reuse Use by or Expiration Date (YYYY-MM-DD) Lot Number Manufacturer's Serial Number Catalog Number or Product Code Attention: See Instructions for Use. <c wa I s Manufacturer Authorized Representative Contains Sufficient for "n" Tests For In Vitro Diagnostic Use Store At or Below Store At or Above Store Between Consult Instructions for Use Fragile, Handle with Care. Keep Dry This end up

Version 1.0

Pub. No. C-344

1DHDT
Lactate Dehydrogenase

INSTRUCTIONS FOR USE


Revision History

Revision History
Date of Revision 2003-10-01 Description of Technical Changes* New format New organization and sections consistent with IVD Directive Materials Required But Not Provided and Sample Dilution - added VITROS 7% BSA Quality Control Material Selection - added statements regarding enzyme activity and ethylene glycol Limitations of the Procedure - removed the statement regarding elevated total protein levels Method Comparison - updated all data and the plot Precision - updated all values References - added all except 1 The change bars indicate the position of a technical amendment to the text with respect to the previous version of the document. Version 1.0

When this Instructions For Use is replaced, sign and date below and retain as specified by local regulations or laboratory policies, as appropriate.

Signature

Obsolete Date

Ortho-Clinical Diagnostics Mandeville House 62 The Broadway Amersham Buckinghamshire HP7 OHJ England Ortho-Clinical Diagnostics, Inc. 100 Indigo Creek Drive Rochester, NY 14626-5101

' Ortho-Clinical Diagnostics


oH company

VITROS is a trademark of Ortho-Clinical Diagnostics, Inc. Ortho-Clinical Diagnostics, Inc., 2003. All rights reserved. Printed in USA.

Pub. No. C-344

Version 1.0

Products

Chemistry

INSTRUCTIONS FOR USE


VITROS Chemistry Products Li DT Slides Intended Use
For in vitro diagnostic use only. VITROS Li DT Slides quantitatively measure lithium (Li) concentration in serum and plasma.

UDT
Lithium

Summary and Explanation of the Test


Lithium is used in the treatment of bipolar (manic-depressive) illness. Lithium measurements are used to monitor patient compliance and therapy and to diagnose potential overdose. Symptoms of lithium intoxication include sluggishness, drowsiness, muscle weakness, and ataxia.'2

Principles of the Procedure


The VITROS Li DT Slide method is performed using the VITROS Li DT Slide and the VITROS Chemistry Products DT Specialty Calibrator Kit on VITROS DT60 II Chemistry Systems. The VITROS Li DT Slide is a multilayered, analytical element coated on a polyester support. A drop of patient sample is deposited on the slide and is evenly distributed by the spreading layer to the underlying layers. The lithium in the sample is specifically bound by the crown-ether azo dye (6-dodecyl-6-(2'-hydroxy5'-(2"-4"-dinitrophenylazo)benzyl)-13, 13-dimethyl-1,4, 8,11-tetraoxacyclotetradecane). As the lithium ion binds to the crownether, a shift in the peak absorbance of the dye occurs. The increase in absorbance is proportional to the concentration of lithium in the sample. The intensity of the dye is measured by reflectance spectrophotometry at the end of incubation.

Reaction Sequence
lithium + crown-ether dye dye complex

Test Type and Conditions


Test Type and Conditions for Li DT
Test Type Colorimetric VITROS DT60 II Module DTSC II Approximate Incubation Time 5 minutes Temperature 37C (98.6F) Wavelength 630 nm Sample Drop Volume 10 uL

Warnings and Precautions


For in vitro diagnostic use only. Take care when handling materials and samples of human origin. Since no test method can offer complete assurance that infectious agents are absent, consider all clinical specimens, controls, and calibrators potentially infectious. Handle specimens, solid and liquid waste, and test components in accordance with local regulations and NCCLS Guideline M293 or other published biohazard safety guidelines. For specific warnings and precautions for calibrators, quality control materials, and other components, refer to the instructions for use for the appropriate VITROS product, or to other manufacturer's product literature.

Version 1.0

Pub. No. C-372

UDT
Lithium

INSTRUCTIONS FOR USE


Reagents

Reagents
Slide Diagram

Slide Ingredients
-- ~~

Reactive ingredients per cm


(6-dodecyl-6-(2'-hydroxy-5'-(2"-4"-dinitrophenylazo)benzyl)-13,13-dimethyl1,4,8,11-tetraoxacyclotetradecane) (crown-ether azo dye) 40 pg.

Other ingredients
Pigment, binders, buffer, surfactants, dye solubilizer and cross-linking agent.
_- - - *

1. Upper slide mount 2. Spreading layer (BaSC>4) buffer, pH 11.0 3. Buffer layer buffer, pH 11.0 4. Reagent layer crowvether azo dye S. Support layer 6. Lower slide mount

Slide Handling
CAUTION; Do not use slides with damaged or incompletely sealed packaging.

Slide Preparation
IMPORTANT: The slide must reach room temperature, 18-28C (64-82F), before the wrapper is opened. Do not use unopened slides that have been at room temperature, 18 -28 C (64-82F) for >48 hours. 1. 2. Remove the unopened slide from the box. Warm the unopened slide for 15 minutes. Unopened slides that remain in the sealed wrapper may be returned to refrigerator or freezer storage. 3. Remove the packaging and place the slide into the loading station within 15 minutes after opening.

Slide Storage and Stability


VITROS Li DT Slides are stable until the expiration date on the carton when they are stored and handled as specified.

Slide Storage and Stability for Li DT Slides Storage Condition


Unopened Room temperature Refrigerated Frozen Room temperature 18-28C(64-82 F) 2-8C (36-46F) <-18C (<0F) 18-28C(64-82 O F)

Opened

Stability <48 hours Until expiration date Until expiration date <15 minutes

Specimen Requirements
WA Handle specimens as biohazardous material.

Specimens Recommended
Serum Plasma: EDTA Heparin Certain collection devices have been reported to affect other analytes and tests." Confirm that your collection devices are compatible with this test.

MPORTANT:

Specimens Not Recommended


Plasma: Fluoride oxalate Lithium heparin

Serum and Plasma


Specimen Collection and Preparation
Collect specimens using standard laboratory procedures.56

Pub. No. C-372

Version 1.0

[3 N/ITFJCp'S

INSTRUCTIONS FOR USE


Testing Procedure Patient Preparation No special patient preparation is necessary. Special Precautions Samples are commonly drawn approximately 12 hours after the last dose of lithium has been taken.7 Centrifuge specimens and remove the serum from the cellular material within 4 hours of collection.8

Li DT
Lithium

Specimen Handling and Storage


WARNING; Handle specimens as biohazardous material. Handle and store specimens in stoppered containers to avoid contamination and evaporation. Mix samples by gentle inversion and bring to room temperature, 18-28C (64-82F), prior to analysis.

Specimen Storage and Stability for Li DT: Serum and Plasma


Storage Room temperature Refrigerated Frozen Temperature 18-28C (64-82F) 2-8C (36-46F) <-18C(<0F) Stability <8 hours <24 hours <6 months

Testing Procedure
Materials Provided
VITROS Chemistry Products Li DT Slides

Materials Required But Not Provided


. VITROS Chemistry Products DT Specialty Calibrator Kit Quality control materials, such as VITROS Chemistry Products DT Control I & II VITROS Chemistry Products 7% BSA Reagent-grade water VITROS DT Pipette

Operating Instructions
I I Refer to the operator's manual for your VITROS DT60 II Chemistry System. Bring all fluids and samples to room temperature, 18-28"C (64-82F), prior to analysis. IMPORTANT:

Sample Dilution
| If lithium concentrations exceed the system's reportable (dynamic) range: 1. Dilute the sample with VITROS 7% BSA or reagent-grade water. 2. Reanalyze. 3. Multiply the results by the dilution factor to obtain an estimate of the original sample's lithium concentration.

Calibration
Required Calibrators
VITROS Chemistry Products DT Specialty Calibrator Kit, bottles 1, 2, and 4

Calibrator Preparation, Handling, and Storage


Refer to the instructions for use for VITROS DT Specialty Calibrator Kit.

Calibration Procedure
Refer to the operator's manual for your VITROS DT60 II Chemistry System.

When to Calibrate
Calibrate: When the slide lot number changes. When critical system parts are replaced due to service or maintenance. When government regulations require. - For example, in the USA, CLIA regulations require calibration or calibration verification at least once every six months.

Version 1.0

Pub. No. C-372

LiDT
Lithium

INSTRUCTIONS FOR USE


Quality Control
The VITROS Li DT test may also need to be calibrated: If quality control results are consistently outside acceptable range. After certain service procedures have been performed. For additional information, refer to the operator's manual for your VITROS DT60 II Chemistry System.

Calculations
Reflectance from the slide is measured at 630 nm after the fixed incubation time. Once a calibration has been performed for each slide lot, lithium concentration in unknown samples can be determined using the software-resident endpoint colorimetric math model and the response obtained from each unknown test slide.

Validity of a Calibration
Calibration parameters are automatically assessed by the VITROS DT60II Chemistry System against a set of quality parameters resident within the analyzer software. Failure to meet any of the pre-defined quality parameters results in a failed calibration. The calibration report should be used in conjunction with quality control results to determine the validity of a calibration.

Reportable (Dynamic) Range Reportable (Dynamic) Range for Li DT


Conventional and SI Units (mmol/L) 0.2-4.0 For out-of-range samples, refer to "Sample Dilution."

Traceability of the Calibration


Values assigned to the VITROS Chemistry Products DT Specialty Calibrator Kit for lithium are traceable to the Certified NIST (National Institute of Standards and Technology) Reference Material, SRM (Standard Reference Material) 924a. The Ortho-Clinical Diagnostics calibration laboratory uses SRM 924a to calibrate the flame atomic absorption spectroscopy method9 to support lithium value assignment for the VITROS DT Specialty Calibrator Kit.

Quality Control
Procedure Recommendations
j WARNING; Handle quality control materials as biohazardous material. Choose control levels that check the clinically relevant range. Analyze quality control materials in the same manner as patient samples, before or during patient sample processing. To verify system performance, analyze control materials: - After calibration. - According to local regulations or at least once each day that the test is being performed. - After specified service procedures are performed. Refer to the operator's manual for your VITROS DT60II System. If control results fall outside your acceptable range, investigate the cause before deciding whether to report patient results. For general quality control recommendations, refer to Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition or other published guidelines. For additional information, refer to the operator's manual for your VITROS DT60 II Chemistry System.

Quality Control Material Selection


I I IMPORTANT: VITROS DT Control I & II are recommended for use with the VITROS DT60 II Chemistry System. Evaluate the performance of other commercial control fluids for compatibility with this test before using for quality control.

Control materials other than VITROS DT Controls I & II may show a difference when compared with other lithium methods if they: - Depart from a true human matrix. - Contain high concentrations of preservatives, stabilizers, or other nonphysiological additives. Do not use control materials stabilized with ethylene glycol.

Quality Control Material Preparation and Storage


Refer to the instructions for use for VITROS Chemistry Products DT Control I & II or to other manufacturer's product literature.

Pub. No. C-372

Version 1.0

[3 VITQCpS

INSTRUCTIONS FOR USE


Expected Values and Reporting Units

UDT
Lithium

Expected Values and Reporting Units


Reference Interval
These serum lithium concentrations are based on the 12-hour standardized concentration, measured on a serum sample obtained 12 hours after the last dose.11 12

Reference Interval for Li DT


Therapeutic 1 3 Toxic 14

Conventional and SI Units (mmol/L) 0.6-1.2 >1.5 >2.5

Potentially Severely

Each laboratory should confirm the validity of these intervals for the population it serves.

Reporting Units and Unit Conversion


The VITROS DT60 II Chemistry System may be programmed to report lithium results in conventional and SI units.

Reporting Units for Li DT

Conventional and SI Units mmol/L

Limitations of the Procedure


Known Interferences
The VITROS Li DT Slide method was screened for interfering substances following NCCLS Protocol EP7.15 The substances listed in the table, when tested at the concentrations indicated, caused the bias shown.

Known Interfering Substances for Li DT


Interferent* Methyl para ben** A/-Acetylcysteine Hemoglobin Interferent Concentration 150mg/dL 180mg/dL 100mg/dL 250 mg/dL 500 mg/dL (10 mmol/L) (11.0 mmol/L) (15.5 umol/L) (38.8 umol/L) (77.5 umol/L) Lithium Concentration Conv./SI Units (mmol/L) 1.0 1.0 1.2 1.2 1.2 Average Bias Conv./SI Units (mmol/L) -0.17 -0.15 +0.04 +0.12 +0.17

**

It is possible that other interfering substances may be encountered. These results are representative; however, your results may differ somewhat due to test-to-test variation. The degree of interference at concentrations other than those listed might not be predictable. A preservative found in some controls, proficiency fluids, and sterile saline flushes

Other Limitations
Certain drugs and clinical conditions are known to alter lithium concentration in vivo. For additional information, refer to one of the published summaries.16'17

Version 1.0

Pub. No. C-372

UDT
Lithium

INSTRUCTIONS FOR USE


Performance Characteristics

Performance Characteristics
Method Comparison

The plot and table show the results of a comparison of samples analyzed on the VITROS DT60 II System with those analyzed using the VITROS 950 System. Testing followed NCCLS Protocol EP9.18

Method Comparison for Li DT: Serum


Conventional and SI Units y =x

Comparative Method: VITROS 950 System (mmol/L)

Method Comparison for Li DT: Serum


Conventional and SI Units (mmol/L) n DT60 II System vs. 950 System 62 Correlation Slope Coefficient 1.01 0.998 Range of Sample Cone. Intercept Sy.x 0.05

0.2-3.9

-0.06

Precision
Precision was evaluated with quality control materials on VITROS the DT60 II System following NCCLS Protocol EP5.19 The data presented are a representation of test performance and are provided as a guideline. Variables such as sample handling and storage, reagent handling and storage, laboratory environment, and system maintenance can affect reproducibility of test results.

Precision for Li DT: Serum


Conventional and SI Units (mmol/L) System VITROS DT60 II Mean Cone. 1.2 2.4 Within Day SD* Within Lab SD** Within Lab CV%** 0.02 0.04 0.04 0.05 3.2 2.1 No. Observ. 84 83 No. Days 21 21

Within Day precision was determined using two runs/day with two replications. Within Lab precision was determined using a single lot of slides and calibrating weekly.

Pub. No. C-372

Version 1.0

g] VITRI

INSTRUCTIONS FOR USE


References

LiDT
Lithium

References
Cade JFJ. Lithium Salts in the Treatment of Psychotic Excitement. Med. J. Aust. 2:349; 1949. Baastrup PC, Schou M. Lithium as a Prophylactic Agent: Its Effect against Recurrent Depressions and Manic-Depressive Psychosis. Arch. Gen. Psychiat. 16:162; 1967. 3. NCCLS. Protection of Laboratory Workers from Instrument Biohazards and Infectious Diseases Transmitted by Blood, Body Fluids and Tissue; Approved Guideline. NCCLS Document M29 (OSBN 1-56238). NCCLS, Wayne, PA 19087; 1997. 4. Calam RR. Specimen Processing Separator Gels: An Update. J Clin Immunoassay. 11:86-90; 1988. 5. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Venipuncture. NCCLS Document H3. Wayne, PA: NCCLS; 1991. 6. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Skin Puncture. NCCLS Document H4. Wayne, PA: NCCLS; 1991. 7. Tietz NW (ed). Fundamentals of Clinical Chemistry, ed. 5. Philadelphia: WB Saunders; 632; 2001. 8. Clinical Laboratory Handbook for Patient Preparation and Specimen Handling. Fascicle IV: Therapeutic Drug Monitoring/Toxicology. Skokie, IL: College of American Pathologists; 1985. 9. Levy AL, Katz EM. A Comparison of Serum Lithium Determinations Using Flame Photometry and Atomic Absorption Spectrophotometry. Clin. Chem. 15:787; 1969. 10. NCCLS. Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition. NCCLS Document C24. Wayne, PA: NCCLS; 1999. 11. Gilman AG et al. Goodman and Gilman's The Pharmacological Basis of Therapeutics, ed. 8. 418-422; 1990. 12. Ellenhorn MJ, Barceloux DG. Medical Toxicology: Diagnosis and Treatment of Human Poisoning. New York: Elsevier; 1042-1045; 1988. 13. Burtis CA, Ashwood ER, (eds) Fundamentals of Clinical Chemistry, ed. 5. Philadelphia: WB Saunders; 1023; 2001. 14. Burtis CA, Ashwood ER, (eds) Fundamentals of Clinical Chemistry, ed. 5. Philadelphia: WB Saunders; 632; 2001. 15. NCCLS. Interference Testing in Clinical Chemistry. NCCLS Document EP7. Wayne, PA: NCCLS; 1986. 16. Young DS. Effects of Drugs on Clinical Laboratory Tests, ed. 4. Washington D.C.: AACC Press; 1995. 17. Friedman RB, Young DS. Effects of Disease on Clinical Laboratory Tests. Washington, D.C.: AACC Press; 1990. 18. NCCLS. Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline. NCCLS Document EP9. Wayne, PA: NCCLS; 1995. 19. NCCLS. User Evaluation of Precision Performance with Clinical Chemistry Devices. NCCLS Document EP5. Wayne, PA: NCCLS; 1992. 1. 2.

Glossary of Symbols
The Glossary of Symbols table defines the symbols used on the VITROS Chemistry Products packaging and on the VITROS Chemistry Products Assay Sheets.

Glossary of Symbols
Do Not Reuse Use by or Expiration Date (YYYY-MM-DD) Lot Number Manufacturer's Serial Number | EC | REP j Manufacturer Authorized Representative Contains Sufficient for "n" Tests For In Vitro Diagnostic Use Store At or Below Store At or Above Store Between Consult Instructions for Use Fragile, Handle with Care. Keep Dry This end up

\2)/

REF

Catalog Number or Product Code Attention: See Instructions for Use.

Version 1.0

Pub. No. C-372

LiDT
Lithium

INSTRUCTIONS FOR USE


Revision History

Revision History
Date of Revision 2003-08-11 Version 1.0 Description of Technical Changes* New format New organization and sections consistent with IVD Directive Specimens Not Recommended - added section Specimen Storage and Stability - updated stability values Materials Required But Not Provided and Sample Dilution - replaced isotonic saline and distilled water with VITROS 7% BSA and reagent-grade water Known Interferences - updated values for hemoglobin Method Comparison - updated all data and the plot Precision - updated all values References - added all except 1, 2,11,12,16,17,18

The change bars indicate the position of a technical amendment to the text with respect to the previous version of the document.

When this Instructions For Use is replaced, sign and date below and retain as specified by local regulations or laboratory policies, as appropriate.

Signature

Obsolete Date

C
I EC REP I Ortho-Clinical Diagnostics Mandeville House 62 The Broadway Amersham Buckinghamshire HP7 OHJ England Ortho-Clinical Diagnostics, Inc. 100 Indigo Creek Drive Rochester, NY 14626-5101

'Ortho-Clinical Diagnostics
ftwm company

VITROS is a trademark of Ortho-Clinical Diagnostics, Inc. Ortho-Clinical Diagnostics, Inc., 2003. All rights reserved. Printed in USA.

Pub. No. C-372

Version 1.0

Chemistry

INSTRUCTIONS FOR USE


VITROS Chemistry Products LIPA DT Slides Intended Use
For in vitro diagnostic use only. VITROS LIPA DT Slides quantitatively measure lipase (LIPA) activity in serum and plasma.

LIPA DT
Lipase

Summary and Explanation of the Test


Lipase is a digestive enzyme that is mainly produced by the acinar cells of the exocrine pancreas. Its physiological role is to hydrolyze the long-chain triglycerides in the small intestine. Serum lipase increases rapidly in patients with acute and recurrent pancreatitis, pancreatic abscess or pseudocyst, pancreatic trauma, pancreatic cancer, common-bile-duct obstruction, and ingestion of drugs that are toxic to the pancreas. It is also increased by most inflammatory conditions in the abdominal cavity, biliary tract disease, abdominal abscesses, and renal failure. Lipase is more specific than total amylase in the diagnosis of acute pancreatitis.1

Principles of the Procedure


The VITROS LIPA DT Slide method is performed using the VITROS LIPA DT Slide and the VITROS Chemistry Products DT Calibrator Kit on VITROS DT60 II Chemistry Systems. The VITROS LIPA DT Slide is a multilayered, analytical element coated on a polyester support. The analysis is based on an enzymatic method described by Mauck.2 A drop of patient sample is deposited on the slide and is evenly distributed by the spreading layer to the underlying layers. The method incorporates colipase which facilitates the adsorption of lipase to the substrate micelles in the presence of bile salts. Lipase then catalyzes the hydrolysis of water-insoluble triacylglycerol esters. The method uses the enzyme diacetinase to convert the substrate 2,3-diacetylglycerol to glycerol. Glycerol kinase converts the glycerol to L-a-glycerophosphate. L-a-glycerophosphate oxidase catalyzes the oxidation of L-a-glycerophosphate to generate hydrogen peroxide. Peroxidase oxidizes a leuco dye to produce a colored dye. The rate of change in reflection density is proportional to the activity of lipase present in the sample.

Reaction Sequence
1 -oleoyl-2,3-diacetylglycerol 2,3-diacetylglycerol glycerol + ATP L-a-glycerophosphate + O2 H2O2 + leuco dye
lipase, colipase pH8.5 diacetinase glycerol kinase MgCI2

2,3-diacetylglycerol + oleic acid glycerol + acetic acid L-a-glycerophosphate + ADP dihydroxyacetone phosphate + H2O2

L- a -glycerophosphate oxidase peroxidase

dye + 2H2O

Test Type and Conditions


Test Type and Conditions for LIPA DT
Test Type Rate VITROS DT60 II Module DTSC II Approximate Incubation Time 5 minutes Temperature 37C (98.6F) Wavelength 540 nm Sample Drop Volume 10 uL

Version 1.0

Pub. No. C-356

LIPADT
Lipase

INSTRUCTIONS FOR USE


Warnings and Precautions

Warnings and Precautions


For in vitro diagnostic use only. Take care when handling materials and samples of human origin. Since no test method can offer complete assurance that infectious agents are absent, consider all clinical specimens, controls, and calibrators potentially infectious. Handle specimens, solid and liquid waste, and test components in accordance with local regulations and NCCLS Guideline M293 or other published biohazard safety guidelines. For specific warnings and precautions for calibrators, quality control materials, and other components, refer to the instructions for use for the appropriate VITROS product, or to other manufacturer's product literature.

Reagents
Slide Diagram

Slide Ingredients
Reactive ingredients per cm2
Diacetinase (Bacillus subtilis, E.C.3.1.1-) 0.54 U; glycerol kinase (. coli or Cellulomonas sp, E.C.2.7.1.30) 0.32 U; L-a-glycerophosphate oxidase {Aerococcus viridans, E.C.1.1.3.21) 0.39 U; peroxidase (horseradish root, E.C.1.11.1.7) 0.62 U; colipase (porcine pancreas) 5.9 U; adenosine triphosphate 0.16 mg; 1-oleoyl-2,3-diacetylglycerol 0.80 mg; and 2-(3,5-dimethoxy4-hydroxyphenyl)-4,5-bis(4-dimethylaminophenyl)imidazo!e (leuco dye) 33 ng.

, 2

3 ^"T ' 4 ~~*~~> "~ "

Other ingredients
Pigment, binders, surfactants, enzyme cofactors, stabilizers, buffer, dye solubilizer, scavenger and cross-linking agent.

1. Upper slide mount 2. Spreading layer {TIO2) colipase 1-oleoyl-2,3-diace1ylglycerol 3. Reagent layer buffer, pH 8.5 diacetinase glycerol kinase ATP L-a-glycerophosphate oxidase peroxidase leuco dye 4. Support layer 5. Lower slide mount

Slide Handling
CAUTION: Do not use slides with damaged or incompletely sealed packaging.

Slide Preparation
IMPORTANT: The slide must reach room temperature, 18-28X: (64-82F), before the wrapper is opened. Do not use unopened slides that have been at room temperature, 18 -28 C (64 -82 F) for >48 hours. 1. 2. Remove the unopened slide from the box. Warm the unopened slide for at least 15 minutes. Unopened slides that remain in the sealed wrapper may be returned to refrigerator or freezer storage. 3. Remove the packaging and place the slide into the loading station within 15 minutes after opening.

Slide Storage and Stability


VITROS LIPA DT Slides are stable until the expiration date on the carton when they are stored and handled as specified.

Slide Storage and Stability for LIPA DT


Slides Unopened

Opened

Storage Condition Room temperature 18-28C (64-82F) Refrigerated 2-8C (36-46F) Frozen <-18C(<0F) Room temperature 18-28C (64-82F)

Stability <48 hours <4 weeks Until expiration date <15 minutes

Pub. No. C-356

Version 1.0

Q VITRCpS

INSTRUCTIONS FOR USE


Specimen Requirements

UPADT
Lipase

Specimen Requirements
I VM Handle specimens as biohazardous material.

Specimens Recommended
Serum Plasma: Heparin Certain collection devices have been reported to affect other analytes and tests4 Confirm that your collection devices are compatible with this test. IMPORTANT:

Specimens Not Recommended


Do not use grossly lipemic specimens. Refer to "Limitations of the Procedure."

Serum and Plasma

Specimen Collection and Preparation


Collect specimens using standard laboratory procedures.5 6 Patient Preparation No special patient preparation is necessary. Special Precautions Ensure equipment is free from soap or giyceroi contamination. Collection tubes with glycerol-lubricated stoppers should not be used. Centrifuge specimens and remove the serum or plasma from the cellular material within 4 hours of collection.7 Specimen Handling and Storage Handle and store specimens in stoppered containers to avoid contamination and evaporation. Mix samples by gentle inversion and bring to room temperature, 18-28C (64-82F), prior to analysis.

Specimen Storage and Stability for LIPA DT: Serum and Plasma7 Storage Temperature Stability
Room temperature Refrigerated Frozen 18-28C(64O-82OF) 2-8C (36-46F) <-18C(<0F) <7 days <3 weeks <5 months

Testing Procedure
Materials Provided
VITROS Chemistry Products LIPA DT Slides

Materials Required But Not Provided


. VITROS Chemistry Products DT Calibrator Kit Quality control materials, such as VITROS Chemistry Products DT Control I & II Isotonic saline Reagent-grade water VITROS DT Pipette

Operating Instructions
Refer to the operator's manual for your VITROS DT60 II Chemistry System. Bring all fluids and samples to room temperature, 18-28C (64-82F), prior to analysis. IMPORTANT;

Sample Dilution
If lipase activities exceed the system's reportable (dynamic) range or if samples are flagged with an L-11 or L-13 error code: 1. Dilute with isotonic saline or reagent-grade water. 2. Reanalyze. 3. Multiply the results by the dilution factor to obtain an estimate of the original sample's lipase activity.

Version 1.0

Pub. No. C-356

1IPADI
Lipase

INSTRUCTIONS FOR USE


Calibration

Calibration
Required Calibrators
VITROS Chemistry Products DT Calibrator Kit, bottles 1, 2, and 4

Calibrator Preparation, Handling, and Storage


Refer to the instructions for use for VITROS DT Calibrator Kit.

Calibration Procedure
Refer to the operator's manual for your VITROS DT60 II Chemistry System.

When to Calibrate
Calibrate: When the slide lot number changes. When critical system parts are replaced due to service or maintenance. When government regulations require. - For example, in the USA, CLIA regulations require calibration or calibration verification at least once every six months. The VITROS LIPA DT test may also need to be calibrated: If quality control results are consistently outside acceptable range. After certain service procedures have been performed. For additional information, refer to the operator's manual for your VITROS DT60 II Chemistry System.

Calculations
Based on sequential readings of the slide's reflectance at 540 nm over the defined incubation period, a rate of change in reflectance is determined. This rate is used in the software-resident multi-point rate calibration model to compute enzyme activity. Once a calibration has been performed for each slide lot, lipase activity in unknown samples can be determined from the rate of change in reflectance measured for each unknown test slide.

Validity of a Calibration
Calibration parameters are automatically assessed by the VITROS DT60 II Chemistry System against a set of quality parameters resident within the analyzer software. Failure to meet any of the pre-defined quality parameters results in a failed calibration. The calibration report should be used in conjunction with quality control results to determine the validity of a calibration.

Reportable (Dynamic) Range Reportable (Dynamic) Range for LIPA DT


Conventional and SI Units (U/L)

10-2000
For out-of-range samples, refer to "Sample Dilution."

Traceability of the Calibration

I
|

Values assigned to the VITROS Chemistry Products DT Calibrator Kit for lipase are traceable to the measurement of lipase activity in a standard triolein emulsion with a pH-Stat analyzer.8

Quality Control
Procedure Recommendations
' ' Handle quality control materials as biohazardous material. Choose control levels that check the clinically relevant range. Analyze quality control materials in the same manner as patient samples, before or during patient sample processing. To verify system performance, analyze control materials: - After calibration. - According to local regulations or at least once each day that the test is being performed. - After specified service procedures are performed. Refer to the operator's manual for your VITROS DT System. If control results fall outside your acceptable range, investigate the cause before deciding whether to report patient results. For general quality control recommendations, refer to Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition9 or other published guidelines. For additional information, refer to the operator's manual for your VITROS DT60 II Chemistry System.

Pub. No. C-356

Version 1.0

El VITRcffS

INSTRUCTIONS FOR USE


Expected Values and Reporting Units

LIPADT
Lipase

Quality Control Material Selection


IMPORTANT; VITROS DT Control I & II are recommended for use with the VITROS DT60/DT60 II Chemistry System. Evaluate the performance of other commercial control fluids for compatibility with this test before using for quality control.

Control materials other than VITROS DT Controls I & II may show a difference when compared with other lipase methods if they: - Depart from a true human matrix. - Contain high concentrations of preservatives, stabilizers, or other nonphysiological additives. Liquid controls are not recommended because they often contain high concentrations of glycerol, which result in L-11 or L-13 codes. Refer to "Limitations of the Procedure." Enzyme activity might also vary with enzyme source, diluent temperature, and activation time during reconstitution. Do not use control materials stabilized with ethylene giycol.

Quality Control Material Preparation and Storage


Refer to the instructions for use for VITROS Chemistry Products DT Control I & II or to other manufacturer's product literature.

Expected Values and Reporting Units


Reference Interval
This reference interval is based on results from an internal study of 496 apparently healthy individuals, ages 10-90. No significant differences in results between male and female populations were observed. Lipase activities tend to increase with age.10

Reference Interval for LIPA DT


Conventional and SI Units (UL) 23-300 Each laboratory should confirm the validity of these intervals for the population it serves.

Reporting Units and Unit Conversion


The VITROS DT60 II Chemistry System may be programmed to report LIPA DT results in conventional and SI units.

Reporting Units for LIPA PT Conventional and SI Units


U/L

Limitations of the Procedure


Known Interferences
Carboxylesterases generally do not interfere with lipase results. Extremely high activity of carboxylesterase (e.g., 51000 U/L) will show a bias of +240 U/L at a lipase activity of 35 U/L. Grossly lipemic samples may show a large negative bias. The VITROS LIPA Slide method was screened for interfering substances following NCCLS Protocol EP7.11 The substances listed in the table, when tested at the concentrations indicated, caused the bias shown.

Known Interfering Substances for LIPA DT


Interferent* 5-Aminosalicylate Interferent Concentration Lipase Activity Conv./SI Units (U/L) 200 Average Bias Conv./SI Units (U/L) -28

23mg/dL

(1.5mmol/L)

It is possible that other interfering substances may be encountered. These results are representative; however, your results may differ somewhat due to test-to-test variation. The degree of interference at concentrations other than those listed might not be predictable.

Other Limitations
Normal endogenous concentrations of glycerol do not interfere with this method; however, samples containing high concentrations of glycerol will be flagged with the L-11 or L-13 error code. Samples flagged with a L-11 or L-13 error code should be diluted and reanalyzed. Refer to "Sample Dilution." Highly elevated glycerol concentrations are usually caused by contamination from rubber stoppers, latex gloves or hyperalimentation fluids. If the L-11 or L-13 error code was caused by glycerol contamination, the final lipase result may be normal. Certain drugs and clinical conditions are known to alter lipase activity in vivo. For additional information, refer to one of the published summaries.1213 Pub. No. C-356

Version 1.0

LIPADT
Lipase

INSTRUCTIONS FOR USE


Performance Characteristics

Performance Characteristics
Methpd Comparison

The plot and table show the results of a comparison of the VITROS DT60 II Chemistry System with the VITROS 700 Chemistry System. Testing followed NCCLS Protocol EP9.14

Method Comparison for LIPA DT: Serum


Conventional and SI Units y =x
2000

1500

5
D to O

1000

500

500

1000

1500

2000

Comparative Method: VITROS 700 System (U/L)

Method Comparison for LIPA DT: Serum


Conventional and SIUnits (U/L) n DT60 II System vs. 700 System 49 Slope 1.00 Correlation Coefficient 0.999 Range of Sample Activity Intercept 0.38 Sy.x 18.62

36-1800

Precision
Precision was evaluated with quality control materials on VITROS DT60 II System following NCCLS Protocol EP5.16 The data presented are a representation of test performance and are provided as a guideline. Variables such as sample handling and storage, reagent handling and storage, laboratory environment, and system maintenance can affect reproducibility of test results.

Precision for LIPA DT: Serum


Conventional and SI Units (U/L) System VITROS DT60 II Mean Activity Within Day SD* Within Lab SD** Within Lab CV%** No. Observ. 88 88 No. Days 22 22 ' 182 1.9 2.7 1.5 5.4 8.2 1.2 674 * Within Day precision was determined using two runs/day with two replications. ** Within Lab precision was determined using a single lot of slides and calibrating weekly.

Pub. No. C-356

Version 1.0

I*J VITRI

INSTRUCTIONS FOR USE


References

LIPADT
Lipase

References
1. 2. 3. 4. 5. 6. 7. Tietz NW (ed). Fundamentals of Clinical Chemistry, ed. 3. Philadelphia: WB Saunders; 399-400; 1987. Mauck JC, Weaver MS, Stanton C. Development of a Kodak Ektachem Clinical Chemistry Slide for Lipase (Abstract). Clin. Chem. 30:1058-1059; 1984. NCCLS. Protection of Laboratory Workers from Instrument Biohazards and Infectious Diseases Transmitted by Blood, Body Fluids and Tissue; Approved Guideline. NCCLS Document M29 (OSBN 1-56238). NCCLS, Wayne, PA 19087; 1997. Calam RR. Specimen Processing Separator Gels: An Update. J Clin Immunoassay. 11:86-90; 1988. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Venipuncture. NCCLS Document H3. Wayne, PA: NCCLS;

1991.
NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Skin Puncture. NCCLS Document H4. Wayne, PA: NCCLS;

1991.
Clinical Laboratory Handbook for Patient Preparation and Specimen Handling. Fascicle VI: Chemistry/Clinical Microscopy. Northfield, IL: College of American Pathologists; 1992. 8. Tietz, NW, Repique, EV. Proposed Standard Method for Measuring Lipase Activity in Serum by a Continuous Sampling Technique. Clin. Chem. 19:1268-1275; 1973. 9. NCCLS. Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition.. NCCLS Document C24. Wayne, PA: NCCLS; 1999. 10. Vankampen L, DiPaola J, Gambino R. Lipase Normals - Some Data. Lab Report 12 (November); 1990. 11. NCCLS. Interference Testing in Clinical Chemistry. NCCLS Document EP7. Wayne, PA: NCCLS; 1986. 12. Young DS. Effects of Drugs on Clinical Laboratory Tests, ed. 4. Washington D C : AACC Press; 1995. 13. Friedman RB, Young DS. Effects of Disease on Clinical Laboratory Tests. Washington, D.C.: AACC Press; 1990. 14. NCCLS. Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline. NCCLS Document EP9. Wayne, PA: NCCLS; 1995. 15. NCCLS. User Evaluation of Precision Performance with Clinical Chemistry Devices. NCCLS Document EP5. Wayne, PA: NCCLS; 1992.

Glossary of Symbols
The Glossary of Symbols table defines the symbols used on the VITROS Chemistry Products packaging and on the VITROS Chemistry Products Assay Sheets.

Glossary of Symbols
Do Not Reuse Use by or Expiration Date (YYYY-MM-DD) Lot Number Manufacturer's Serial Number Manufacturer Authorized Representative Contains Sufficient for "n" Tests For In Vitro Diagnostic Use Store Between Consult Instructions for Use Fragile, Handle with Care. Keep Dry This end up

REF

Catalog Number or Product Code Attention: See Instructions for Use.

X I

Store At or Below Store At or Above

Version 1.0

Pub. No. C-356

LIPflDT
Lipase

INSTRUCTIONS FOR USE


Revision History

Revision History
Date of Revision 2003-03-28 Version 1.0 Description of Technical Changes* New organization and sections consistent with IVD Directive Specimens Not Recommended - plasma: removed oxalate/fluoride, citrate, and EDTA Specimen Storage and Stability - updated all stabilities Quality Control Material Selection - added statements regarding liquid controls, enzyme activity, and ethylene glycol Limitations of the Procedure - removed sodium deoxycholate; updated bias due to carboxylesterase; added L-13 error code and removed >AR Method Comparison - corrected the slope value Precision - updated all values References - added all but 10

The change bars indicate the position of a technical amendment to the text with respect to the previous version of the document.

When this Instructions For Use is replaced, sign and date below and retain as specified by local regulations or laboratory policies, as appropriate.

Signature

Obsolete Date

C
Ortho-Clinical Diagnostics Mandeville House 62 The Broadway Amersham Buckinghamshire HP7 OHJ England Ortho-Clinical Diagnostics, Inc. 100 Indigo Creek Drive Rochester, NY 14626-5101

'Ortho-Clinical Diagnostics
^c&mcH company

VITROS is a trademark of Ortho-Clinical Diagnostics, Inc. Ortho-Clinical Diagnostics, Inc., 2003. All rights reserved. Printed in USA.

Pub. No. C-356

Version 1.0

Products

Chemistry

INSTRUCTIONS FOR USE


VITROS Chemistry Products Mg DT Slides Intended Use
For in vitro diagnostic use only. VITROS Mg DT Slides quantitatively measure magnesium (Mg) concentration in serum and plasma.

MgDT
Magnesium

Summary and Explanation of the Test


Magnesium is predominantly an intracellular cation and is essential in enzyme reactions. Magnesium deficiency may cause weakness, tremors, tetany, and convulsions. Hypomagnesemia is associated with hypocalcemia, alcoholism, some types of malnutrition, malabsorption, chronic hemodialysis, and pregnancy. Increased serum magnesium concentrations occur in patients with renal failure, dehydration, and Addison's disease.1

Principles of the Procedure


The VITROS Mg DT Slide method is performed using the VITROS Mg DT Slide and the VITROS Chemistry Products DT Calibrator Kit on VITROS DT60/DT60 II Chemistry Systems. The VITROS Mg DT Slide is a multilayered, analytical element coated on a polyester support. A drop of patient sample is deposited on the slide and is evenly distributed by the spreading layer to the underlying layers. Magnesium (both free and protein-bound) from the sample then reacts with the formazan dye derivative in the reagent layer; the high magnesium affinity of the dye dissociates magnesium from binding proteins. The resulting magnesium-dye complex causes a shift in the dye absorption maximum. The amount of dye complex formed is proportional to the magnesium concentration present in the sample and is measured by reflection density.

Reaction Sequence
Mg2 + Ca+2 Mg+2 + formazan dye derivative Mg*2 + Ca*2-chelator complex
pH 9.75

Mg -dye complex

Test Type and Conditions


Test Type and Conditions for Mg DT
Test Type Colorimetric VITROS DT60/DT60 II Module DT60/DT60 II Approximate Incubation Time 5 minutes Temperature 37C (98.6F) Wavelength 660 nm Sample Drop Volume 10 uL

Warnings and Precautions


For in vitro diagnostic use only. Take care when handling materials and samples of human origin. Since no test method can offer complete assurance that infectious agents are absent, consider all clinical specimens, controls, and calibrators potentially infectious. Handle specimens, solid and liquid waste, and test components in accordance with local regulations and NCCLS Guideline M29Z or other published biohazard safety guidelines. For specific warnings and precautions for calibrators, quality control materials, and other components, refer to the instructions for use for the appropriate VITROS product, or to other manufacturer's product literature.

Version 1.0

Pub. No. C-349

VITFJCpS gj

NlgDT
Magnesium

INSTRUCTIONS FOR USE


Reagents

Reagents
Slide Diagram

Slide Ingredients
Reactive ingredients per cm
1,2-bis(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (calcium chelator) 242 ( and 1,5-bis(2-hydroxy-3,5-dichlorophenyl)-3-cyanoformazan (dye) 38 \xg.

Other ingredients
Pigment, binders, buffer, dye solubilizer, surfactants, cross-linking agent and stabilizer.

4 5

1. Upper slide mount 2. Spreading layer (TiO2) 3. Reagent layer calcium chelator buffer, pH 9.75 formazan dye 4. Support layer 5. Lower slide mount

Slide Handling
Do not use slides with damaged or incompletely sealed packaging.

Slide Preparation
IMPORTANT: The slide must reach room temperature, 18-28X: (64-82F), before the wrapper is opened. Do not use unopened slides that have been at room temperature, 18 -28 C (64 -82 F) for >48 hours. 1. 2. Remove the unopened slide from the box. Warm the unopened slide for 15 minutes. Unopened slides that remain in the sealed wrapper may be returned to refrigerator or freezer storage. 3. Remove the packaging and place the slide into the loading station within 15 minutes after opening.

Slide Storage and Stability


VITROS Mg OT Slides are stable until the expiration date on the carton when they are stored and handled as specified.

Slide Storage and Stability for Mg DT Slides Storage Condition


Unopened Opened Room temperature Frozen Room temperature 18-28C (64-82F) <-18C (<0F) 18-28C (64-82F)

Stability <48 hours Until expiration date <15 minutes

Specimen Requirements
Handle specimens as biohazardous material. CAUTION: Protective gloves manufactured with magnesium stearate (talc) powders may cause elevated test results because of the contamination of sample handling supplies (for example, pipette tips, transfer pipettes, sample cups and caps). Supplies that have come in contact with powdered gloves may subsequently contaminate the test specimen during sample metering. Gloves labeled as "powder-free" may contain some contaminating powder agents on the inside of the glove.

NOTE:

Specimens Recommended
Serum Plasma:3 Heparin Certain collection devices have been reported to affect other analytes and tests.4 Confirm that your collection devices are compatible with this test.

IMPORTANT:

Pub. No. C-349

Version 1.0

INSTRUCTIONS FOR USE


Testing Procedure Specimens Not Recommended
. Plasma:5 EDTA Fluoride oxalate Citrate

Mg OT
Magnesium

Serum and Plasma


Specimen Collection and Preparation
Collect specimens using standard laboratory procedures.6 7 Patient Preparation No special patient preparation is necessary. Special Precautions Hemolyzed specimens can cause falsely elevated results due to intracellular magnesium levels.8 Centrifuge specimens and remove the serum or plasma from the cellular material as soon as possible after collection.1 Specimen Handling and Storage Handle specimens as biohazardous material. Handle and store specimens in stoppered containers to avoid contamination and evaporation. Mix samples by gentle inversion and bring to room temperature, 18-28C (64-82F), prior to analysis. Specimen Storage and Stability for Mg DT: Serum and Plasma5 Storage Temperature Stability Room temperature 18-28C(64-82''F) <1 week Refrigerated 2-8C (36-46F) <1 week Frozen <-18C(<0F) <1 month

Testing Procedure
Materials Provided
VITROS Chemistry Products Mg DT Slides

Materials Required But Not Provided


VITROS Chemistry Products DT Calibrator Kit Quality control materials, such as VITROS Chemistry Products DT Control I & II Isotonic saline Reagent-grade water VITROS DT Pipette

Operating Instructions
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System. Bring all fluids and samples to room temperature, 18-28C (64-82F), prior to analysis. IMPORTANT:

Sample Dilution
If magnesium concentrations exceed the system's reportable (dynamic) range: 1. Dilute the sample with isotonic saline or reagent-grade water. 2. Reanalyze. 3. Multiply the results by the dilution factor to obtain an estimate of the original sample's magnesium concentration.

Version 1.0

Pub. No. C-349

MgDT
Magnesium

INSTRUCTIONS FOR USE


Calibration

Calibration
Required Calibrators
VITROS Chemistry Products DT Calibrator Kit, bottles 1,2, and 3

Calibrator Preparation, Handling, and Storage


Refer to the instructions for use for VITROS DT Calibrator Kit.

Calibration Procedure
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

When to Calibrate
Calibrate: When the slide lot number changes. When critical system parts are replaced due to service or maintenance. When government regulations require. - For example, in the USA, CLIA regulations require calibration or calibration verification at least once every six months. The VITROS Mg DT test may also need to be calibrated: If quality control results are consistently outside acceptable range. After certain service procedures have been performed. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Calculations

Reflectance from the slide is measured at 660 nm after the fixed incubation time. Once a calibration has been performed for each slide lot, magnesium concentration in unknown samples can be determined using the software-resident endpoint colorimetric math model and the response obtained from each unknown test slide.

Validity of a Calibration
Calibration parameters are automatically assessed by the VITROS DT60/DT60 ll.Chemistry System against a set of quality parameters resident within the analyzer software. Failure to meet any of the pre-defined quality parameters results in a failed calibration. The calibration report should be used in conjunction with quality control results to determine the validity of a calibration.

Reportable (Dynamic) Range Reportable (Dynamic) Range for Mg DT


Conventional (mgfdL) SI Units (mmol/L) 0.1-2.9

0.2-7.0

For out-of-range samples, refer to "Sample Dilution."

Traceability of the Calibration

I
|

Values assigned to the VITROS Chemistry Products DT Calibrator Kit for magnesium are traceable to the Certified

NIST (National Institute of Standards and Technology) Reference Material, SRM (Standard Reference Material) 929. The Ortho-Clinical Diagnostics calibration laboratory uses SRM" 929 to calibrate the flame atomic absorption spectroscopy method9 to support magnesium value assignment for the VITROS DT Calibrator Kit.

Quality Control
Procedure Recommendations
':' Handle quality control materials as biohazardous material. Choose control levels that check the clinically relevant range. Analyze quality control materials in the same manner as patient samples, before or during patient sample processing. T o verify system performance, analyze control materials: After calibration. According to local regulations or at least once each day that the test is being performed. After specified service procedures are performed. Refer to the operator's manual for your VITROS DT60/DT60II System.

If control results fall outside your acceptable range, investigate the cause before deciding whether to report patient results.

Pub. No. C-349

Version 1.0

INSTRUCTIONS FOR USE


Expected Values and Reporting Units

NlgDT
Magnesium

For general quality control recommendations, refer to Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition or other published guidelines. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Quality Control Material Selection


IMPORTANT: VITROS DT Control I & II are recommended for use with the VITROS DT60/DT60 II Chemistry System. Evaluate the performance of other commercial control fluids for compatibility with this test before using for quality control.

Control materials other than VITROS DT Controls I & II may show a difference when compared with other magnesium methods if they: - Depart from a true human matrix. - Contain high concentrations of preservatives, stabilizers, or other nonphysiological additives. Do not use control materials stabilized with ethylene glycol.

Quality Control Material Preparation and Storage


Refer to the instructions for use for VITROS Chemistry Products DT Control I & II or to other manufacturer's product literature.

Expected Values and Reporting Units


Reference Interval
This reference interval is the central 95% of results from an internal study of 288 apparently healthy individuals from a working population (58 females and 230 males).

Reference Interval for Mg DT


Conventional Units (mg/dL)

1.6-2.3

SI Units (mmol/L) 0.7-1.0

Each laboratory should confirm the validity of these intervals for the population it serves.

Reporting Units and Unit Conversion


The VITROS DT60/DT60 II Chemistry System may be programmed to report magnesium results in conventional and SI units.

Reporting Units and Unit Conversion for Mg DT Conventional Units SI Units


mg/dL mmol/L (mg/dL x 0.4114)

Limitations of the Procedure


Known Interferences
The VITROS Mg DT Slide method was screened for interfering substances following NCCLS Protocol EP7." The substances listed in the table, when tested at the concentrations indicated, caused the bias shown.

Known Interfering Substances for Mg DT


Interferent* Calcium Interferent Concentration 20 mg/dL (5.0 mmol/L) Magnesium Concentration Conv. (mg/dL) SI (mmol/L) 2.3 0.9 Average Bias Conv. (mg/dL) SI (mmol/L) +0.3 +0.12

Other Limitations
Certain drugs and clinical conditions are known to alter magnesium concentrations in vivo. For additional information, refer to one of the published summaries.1213

Version 1.0

Pub. No. C-349

VITFJLpS Q

NlgDT
Magnesium

INSTRUCTIONS FOR USE


Performance Characteristics

Performance Characteristics
Method Comparison

The plots and table show the results of a comparison of samples analyzed on the VITROS DT60 II System with those analyzed using the VITROS 950 System. Testing followed NCCLS Protocol EP9.14 Method Comparison for Mg DT: Serum Conventional Units SI Units

y-x

I
o t

10

w O

Comparative Method: VITROS 950 System (mg/dL)

Comparative Method: VITROS 950 System (mmol/L)

Method Comparison for Mg DT: Serum


Correlation Slope Coefficient 0.98 0.998 Conventional Units (mg/dL) Range of Sample Cone. Intercept Sy.x SI Units (mmol/L) Range of Sample Cone. 0.14-2.85 Intercept 0.05 Sy.x 0.06

n DT60 II Sys.tem vs. 950 System 63

0.3-6.9

0.11

0.14

Precision
Precision was evaluated with quality control materials on the VITROS DT60 II System following NCCLS Protocol EP5.15 The data presented are a representation of test performance and are provided as a guideline. Variables such as sample handling and storage, reagent handling and storage, laboratory environment, and system maintenance can affect reproducibility of test results.

Precision for Mg DT: Serum


Conventional Units (mg/dL) System VITROS DT60 II Mean Cone. 2.3 5.0 Within Day SD* 0.05 0.08 Within Lab SD** 0.06 0.12 Mean Cone. 1.0 2.0 SI Units (mmol/L) Within Within Day SD* Lab SD** 0.02 0.02 0.03 0.05 Within Lab CV%** 2.5 2.4 No. Observ. 88 88 No. Days 22 22

Within Day precision was determined using two runs/day with two replications. Within Lab precision was determined using a single lot of slides and calibrating weekly.

Pub.

No. C-349

Version 1.0

INSTRUCTIONS FOR USE


References

MgDT
Magnesium

References
1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. Tietz NW(ed). Fundamentals of Clinical Chemistry, ed. 3. Philadelphia: WB Saunders; 521-524; 1987. NCCLS. Protection of Laboratory Workers from Instrument Biohazards and Infectious Diseases Transmitted by Blood, Body Fluids and Tissue; Approved Guideline. NCCLS Document M29 (OSBN 1-56238). NCCLS, Wayne, PA 19087; 1997. Doumas BT, et al. Differences Between Values for Plasma and Serum in Tests Performed in the Ektachem 700 XR Analyzer, and Evaluation of "Plasma Separator Tubes (PST)" Clin. Chem. 35:151-153; 1989. Calam RR. Specimen Processing Separator Gels: An Update. J Clin Immunoassay. 11:86-90; 1988. Clinical Laboratory Handbook for Patient Preparation and Specimen Handling. Fascicle VI: Chemistry/Clinical Microscopy. Northfield, IL: College of American Pathologists; 1992. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Venipuncture. NCCLS Document H3. Wayne, PA: NCCLS; 1991. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Skin Puncture. NCCLS Document H4. Wayne, PA: NCCLS;

1991.
Young DS. Effects of Preanalytical Variables on Clinical Laboratory Tests. Washington D.C.: AACC Press; 4-120; 1993. Kaplan L, Pesce A. Clinical Chemistry: Theory, Analysis, and Correlation. CV Mosby; 1069; 1984. NCCLS. Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition. NCCLS Document C24. Wayne, PA: NCCLS; 1999. NCCLS. Interference Testing in Clinical Chemistry. NCCLS Document EP7. Wayne, PA: NCCLS; 1986. Young DS. Effects of Drugs on Clinical Laboratory Tests, ed. 4. Washington D.C.: AACC Press; 1995. Friedman RB. Young DS. Effects of Disease on Clinical Laboratory Tests. Washington, D C : AACC Press; 1990. NCCLS. Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline. NCCLS Document EP9. Wayne, PA: NCCLS; 1995. NCCLS. User Evaluation of Precision Performance with Clinical Chemistry Devices. NCCLS Document EP5. Wayne, PA: NCCLS; 1992.

Glossary of Symbols
The Glossary of Symbols table defines the symbols used on the VITROS Chemistry Products packaging and on the VITROS Chemistry Products Assay Sheets.

Glossary of Symbols
Do Not Reuse Use by or Expiration Date (YYYY-MM-DD) Lot Number Manufacturer's Serial Number Catalog Number or Product Code Attention: See Instructions for Use. Manufacturer Authorized Representative Contains Sufficient for "n" Tests For In Vitro Diagnostic Use Store At or Below Store At or Above Store Between Consult Instructions for Use Fragile, Handle with Care. Keep Dry This end up

Version 1.0

Pub.

No. C-349

NlgDT
Magnesium

INSTRUCTIONS FOR USE


Revision History

Revision History
Date of Revision 2003-10-01 Description of Technical Changes* > New format > New organization and sections consistent with IVD Directive > Specimens Recommended - plasma: updated to heparin > Limitations of the Procedure-removed inorganic phosphorous > Method Comparison -updated all comparisons and plots Precision: updated values References - added all except 6, 8 The change bars indicate the position of a technical amendment to the text with respect to the previous version of the document. Version 1.0

When this Instructions For Use is replaced, sign and date below and retain as specified by local regulations or laboratory policies, as appropriate.

Signature

Obsolete Date

C
Ortho-Clinical Diagnostics Mandeville House 62 The Broadway Amersham Buckinghamshire HP7 OHJ England Ortho-Clinical Diagnostics, Inc. 100 Indigo Creek Drive Rochester, NY 14626-5101

' Ortho-Clinical Diagnostics


H company

VITROS is a trademark of Ortho-Clinical Diagnostics, Inc. Ortho-Clinical Diagnostics, Inc., 2003. All rights reserved. Printed in USA.

Pub.

No. C-349

Version 1.0

Chemistry

VITROS Chemistry Products Na+ DT Slides

INSTRUCTIONS FOR USE

Na DT
Sodium

Intended Use
For in vitro diagnostic use only. VITROS Na+ DT Slides quantitatively measure sodium (Na+) concentration in serum and plasma.

Summary and Explanation of the Test


Sodium is the major cation of extracellular fluids. The kidneys regulate sodium content of the body. Low sodium levels may be caused by excessive urine loss, diarrhea, Addison's disease, and renal tubular disease. High sodium levels may occur in severe dehydration, some types of brain injury, diabetic coma, and excessive intake of sodium salts.1

Principles of the Procedure


The VITROS Na+ DT Slide method is performed using the VITROS Na+ DT Slide and the VITROS Chemistry Products DT Calibrator Kit on VITROS DT60/DT60 II Chemistry Systems. The VITROS Na* DT Slide is a multilayered, analytical element coated on a polyester support that uses direct potentiometry2 for measurement of sodium ions. The slide consists of two ion-selective electrodes, each containing methyl monensin (an ionophore for sodium), a reference layer, and a silver layer and a silver chloride layer coated on a polyester support. A drop of patient sample and a drop of VITROS DT Reference Fluid on separate halves of the slide results in migration of both fluids toward the center of the paper bridge. A stable liquid junction is formed that connects the reference electrode to the sample electrode. Each electrode produces an electrochemical potential in response to the activity of sodium. The potential difference between the two electrodes is proportional to the sodium concentration in the sample.

Test Type and Conditions


Test Type and Conditions for Na* DT
Test Type Potentiometric VITROS DT60/DT60 II Module DTE/DTE II Approximate Incubation Time 90 or 180 seconds* Temperature 25C (77F) Drop Volume Sample: Reference 10 uL Fluid: 10 uL

Assay time is determined by the Calibration Data Module (CDM).

Warnings and Precautions


For in vitro diagnostic use only. Take care when handling materials and samples of human origin. Since no test method can offer complete assurance that infectious agents are absent, consider all clinical specimens, controls, and calibrators potentially infectious. Handle specimens, solid and liquid waste, and test components in accordance with local regulations and NCCLS Guideline M293 or other published biohazard safety guidelines. For specific warnings and precautions for calibrators, quality control materials, and other components, refer to the instructions for use for the appropriate VITROS product, or to other manufacturer's product literature.

Version 1.0

Pub. No. C-307

NaDT
Sodium

INSTRUCTIONS FOR USE


Reagents

Reagents
Slide Diagram

Slide Ingredients Reactive ingredients per cm


2

___ 1

Silver 0.4 mg; silver chloride 0.2 mg; sodium chloride 0.3 mg and methyl monensin 50 ng.

* _- 3 _ - *

Other ingredients
Binders, buffer, plasticizers, stabilizer, surfactants and nickel.

"'Y ~ .
7

1. Upper slide mount 2. Paper Bridge 3. Ion-selective membrane Methyl monensin 4. Reference layer NaCI Buffer, at pM 5.6 5. Silver, silver chloride layer 6. Support layer 7. Lower slide mount

Slide Handling
CAUTION: Do not use slides with damaged or incompletely sealed packaging.

Slide Preparation
IMPORTANT: The slide must reach room temperature, 18-28 C (64 -82 f), before the wrapper is opened. Do not use unopened slides that have been at room temperature, 18 -28 C (64 -82 F) for >48 hours. 1. Remove the unopened slide from the box. 2. Warm the unopened slide for 15 minutes. Unopened slides that remain in the sealed wrapper may be returned to refrigerator or freezer storage. 3. Remove the packaging and place the slide into the loading station within 15 minutes after opening.

Slide Storage and Stability


VITROS Na* DT Slides are stable until the expiration date on the carton when they are stored and handled as specified.

Slide Storage and Stability for Na* DT


Slides Unopened

Opened

Storage Condition Room temperature 18-28C (64-82F) Refrigerated 2-8C (36-46F) Frozen <-18C (<0F) Room temperature 18-28C (64-82F)

Stability <48 hours Until expiration date Until expiration date <15 minutes

Specimen Requirements
Handle specimens as biohazardous material. Specimens Recommended
Serum Plasma:4 Heparin (Sodium heparin will increase the measured sodium value by approximately 0.5 mmol/L.) Certain collection devices have been reported to affect other analytes and tests.5 Confirm that your collection devices are compatible with this test.

IMPORTANT:

Serum and Plasma Specimen Collection and Preparation


Collect specimens using standard laboratory procedures.6'7 Patient Preparation No special patient preparation is necessary.

Pub. No. C-307

Version 1.0

INSTRUCTIONS FOR USE


Testing Procedure
Special Precautions Do not draw specimen from an arm receiving an intravenous transfusion. Fibrin clots may cause incomplete sampling of the specimen.6 - Allow specimens to clot completely in order to prevent fibrin clots. - Inspect plasma specimens for fibrin clots. Centrifuge specimens and remove the serum from the cellular material within 2 days of collection.9 Specimen Handling and Storage Handle and store specimens in stoppered containers to avoid contamination and evaporation. Mix samples by gentle inversion and bring to room temperature, 18-28C (64-82F), prior to analysis.

NaDT
Sodium

Specimen Storage and Stability for Na* DT: Serum and Plasma9
Storage Room temperature Refrigerated Frozen Temperature 18O-28C(64O-82F) 2-8C (36-46F) <-18C(<0F) Stability <4 days <1 week <6 months

Testing Procedure
Materials Provided
VITROS Chemistry Products Na* DT Slides

Materials Required But Not Provided


. VITROS Chemistry Products DT Calibrator Kit Quality control materials, such as VITROS Chemistry Products DT Control I & II VITROS Chemistry Products DT Reference Fluid VITROS DTE Dual Sample Cups VITROS DTE Pipette

Operating Instructions
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System. Bring all fluids and samples to room temperature, 18-28C (64-82F), prior to analysis. IMPORTANT:

Sample Dilution
Sodium concentrations outside the reportable (dynamic) range are not expected. Diluted samples should not be analyzed with VITROS Na* DT Slides because dilution changes both the concentration of solids in plasma water and the ionic strength of the sample.

Calibration
Required Calibrators
VITROS Chemistry Products DT Calibrator Kit, bottles 1 and 2

Calibrator Preparation, Handling, and Storage


Refer to the instructions for use for VITROS DT Calibrator Kit.

Calibration Procedure
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System. MOTE: Calibrate sodium in duplicate by running each bottle twice.

When to Calibrate
Calibrate: When the slide lot number changes. When the VITROS DT Reference Fluid lot number changes. When critical system parts are replaced due to service or maintenance. When government regulations require. - For example, in the USA, CLIA regulations require calibration or calibration verification at least once every six months. Version 1.0 Pub. No. C-307 3

Na DT
Sodium

INSTRUCTIONS FOR USE


Quality Control
The VITROS Na* DT test may also need to be calibrated: If quality control results are consistently outside acceptable range. After certain service procedures have been performed. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Calculations
The VITROS DT60/DT60 II Chemistry System measures the potential difference in millivolts between the two electrodes of a potentiometric slideone in contact with the sample to be analyzed and the other in contact with the electrolyte reference fluid. A linear relationship exists between the measured potential difference observed on the slide and the logarithm of sodium concentration, i.e. the Nernst equation for ion-selective electrodes. Once the calibration has been established for each slide lot, unknown sodium concentrations for a given sample can be determined using the software-resident math model and the measured potential difference.

Validity of a Calibration
Calibration parameters are automatically assessed by the VITROS DT60/DT60 II Chemistry System against a set of quality parameters resident within the analyzer software. Failure to meet any of the pre-defined quality parameters results in a failed calibration. The calibration report should be used in conjunction with quality control results to determine the validity of a calibration.

Reportable (Dynamic) Range Reportable (Dynamic) Range for Na* PT


Conventional and SI Units (mmol/L)

95-215
For out-of-range samples, refer to "Sample Dilution."

Traceability of the Calibration


Values assigned to the VITROS Chemistry Products DT Calibrator Kit for sodium are traceable to the Certified NIST (National Institute of Standards and Technology) Reference Material, SRM" (Standard Reference Material) 919a. The Ortho-Clinical Diagnostics calibration laboratory uses SRM" 919a to calibrate the flame atomic emission spectroscopy method10 to support sodium value assignment for the VITROS DT Calibrator Kit.

Quality Control
Procedure Recommendations
| Handle quality control materials as biohazardous material. Choose control levels that check the clinically relevant range. Analyze quality control materials in the same manner as patient samples, before or during patient sample processing. To verify system performance, analyze control materials: - After calibration. - According to local regulations or at least once each day that the test is being performed. - After specified service procedures are performed. Refer to the operator's manual for your VITROS DT60/DT60 II System. If control results fall outside your acceptable range, investigate the cause before deciding whether to report patient results. For general quality control recommendations, refer to Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition'1'' or other published guidelines. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Quality Control Material Selection


I IMPORTANT; VITROS DT Control I & II are recommended for use with the VITROS DT60/DT60 II Chemistry System. Evaluate the performance of other commercial control fluids for compatibility with this test before using for quality control.

Control materials other than VITROS DT Controls I & II may show a difference when compared with other sodium methods if they: - Depart from a true human matrix. - Contain high concentrations of preservatives, stabilizers, or other nonphysiological additives. Do not use control materials stabilized with ethylene glycol.

Pub.

No. C-307

Version 1.0

INSTRUCTIONS FOR USE


Expected Values and Reporting Units

NaDT
Sodium

Quality Control Material Preparation and Storage


Refer to the instructions for use for VITROS Chemistry Products DT Control I & II or to other manufacturer's product literature.

Expected Values and Reporting Units


I R e f e r e n c e Interval The serum reference interval is the central 95% of results from an internal study of 60 apparently healthy adults from a working population. No significant differences between results from the male and female populations were observed.

Reference Interval for Na+ DT


Conventional and SI Units (mmol/L) 137-145 Each laboratory should confirm the validity of these intervals for the population it serves.

Reporting Units
The VITROS DT60/DT60 II Chemistry System may be programmed to report sodium results in conventional and SI units.

Reporting Units for Na* DT


Conventional and SI Units mmol/L

Limitations of the Procedure


Known Interferences
Specimens contaminated with cationic surfactants show a positive interference (e.g., benzalkonium chloride at 10 mg/L caused a 50 mmol/L apparent increase in sodium). NOTE: Heparinized catheters may contain benzalkonium chloride. Specimens drawn through these catheters should not be used.

Other Limitations
Certain drugs and clinical conditions are known to alter sodium concentration in vivo. For additional information, refer to one of the published summaries.12'13

Version 1.0

Pub. No. C-307

NaDT
Sodium

INSTRUCTIONS FOR USE


Performance Characteristics

Performance Characteristics
Method Comparison
The plots and table show the results of a comparison of samples analyzed on the VITROS DT60 II System with those analyzed using the Flame Photometry comparative method.10 Testing followed NCCLS Protocol EP9." The table also shows the results of a comparison of the VITROS Na* DT Slide at an assay time of approximately 90 seconds with the VITROS Na+ DT Slide at an assay time of approximately 180 seconds.

Method Comparison for Na* DT: Serum


Conventional and SI Units

250

200

150

100

50 50
100

150

200

250

Comparative Method: Flame Photometry (mmol/L)

Method Comparison for Na *DT: Serum


Conventional and SIUnits (mmol/L) n DT60II System vs. comparative method 90 seconds vs. 180 seconds 58 59 Slope 0.99 0.98 Correlation Coefficient 0.960 0.999 Range of Sample Cone. Intercept -1.43 +2.38 Sy.x 2.97 0.85

117-166 101-185

Precision
Precision was evaluated with quality control materials on the VITROS DT60 II System following NCCLS Protocol EP5.'6 The data presented are a representation of test performance and are provided as a guideline. Variables such as sample handling and storage, reagent handling and storage, laboratory environment, and system maintenance can affect reproducibility of test results.

Precision for Na* DT: Serum


Conventional and SI Units (mmol/L) System VITROS DT60 II Mean Cone . 115 133 Within Day SD* 0.7 0.8 Within Lab SD** Within Lab CV%** 1.3 1.4 1.2 1.0 No. Observ. 87 88 No. Days 22 22

Within Day precision was determined using two runs/day with two replications. Within Lab precision was determined using a single lot of slides and calibrating weekly.

Pub. No. C-307

Version 1.0

INSTRUCTIONS FOR USE


References

Na DT
Sodium

References
1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. Tietz NW(ed). Fundamentals of Clinical Chemistry, ed. 3. Philadelphia: WB Saunders; 614-616; 1987. Siggard-Anderson O. Electrochemistry, in Tietz NW (ed). Textbook of Clinical Chemistry. Philadelphia: WB Saunders; 110-125; 1986. NCCLS. Protection of Laboratory Workers from Instrument Biohazards and Infectious Diseases Transmitted by Blood, Body Fluids and Tissue; Approved Guideline. NCCLS Document M29 (OSBN 1-56238). NCCLS, Wayne, PA 19087; 1997. Doumas BT, et al. Differences Between Values for Plasma and Serum in Tests Performed in the Ektachem 700 XR Analyzer, and Evaluation of "Plasma Separator Tubes (PST)." Clin. Chem. 35:151-153; 1989. Calam RR. Specimen Processing Separator Gels: An Update. J Clin Immunoassay. 11:86-90; 1988. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Venipuncture. NCCLS Document H3. Wayne, PA: NCCLS;

1991.
NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Skin Puncture. NCCLS Document H4. Wayne, PA: NCCLS; 1991. Slockbower JM, BlumenfeldTA (ed.). Collection and Handling of Laboratory Specimens. Philadelphia: Lippincott Co; 201; 1983. Clinical Laboratory Handbook for Patient Preparation and Specimen Handling. Fascicle VI: Chemistry/Clinical Microscopy. Northfield, IL: College of American Pathologists; 1992. Velapoldi R.A., et al. A reference method for the determination of sodium in serum. National Institute of Standards and Technology Special Publication 260-60, Gaithersburg. MD, 1978. NCCLS. Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition. NCCLS Document C24. Wayne, PA: NCCLS; 1999. Young DS. Effects of Drugs on Clinical Laboratory Tests, ed. 4. Washington D.C.: AACC Press; 1995. Friedman RB, Young DS. Effects of Disease on Clinical Laboratory Tests. Washington, D.C.: AACC Press; 1990. NCCLS. Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline. NCCLS Document EP9. Wayne, PA: NCCLS; 1995. NCCLS. User Evaluation of Precision Performance with Clinical Chemistry Devices. NCCLS Document EP5. Wayne, PA: NCCLS; 1992.

Glossary of Symbols
The Glossary of Symbols table defines the symbols used on the VITROS Chemistry Products packaging and on the VITROS Chemistry Products Assay Sheets.

Glossary of Symbols
Do Not Reuse Use by or Expiration Date (YYYY-MM-DD) Lot Number Manufacturer's Serial Number \/ BEP | Manufacturer Authorized Representative Contains Sufficient for "n" Tests For In Vitro Diagnostic Use Store At or Below Store At or Above Store Between Consult Instructions for Use Fragile, Handle with Care. Keep Dry This end up

REJF

Catalog Number or Product Code Attention: See Instructions for Use.

Version 1.0

Pub. No. C-307

Ha DT
Sodium

INSTRUCTIONS FOR USE


Revision History

Revision History
Date of Revision 2003-10-01 Version 1.0 Description of Technical Changes* > New format > New organization and sections consistent with IVD Directive Reference Interval - corrected the sample size > Limitations of the Procedure - removed the statement regarding ethanol > Method Comparison - updated all data and the plot > Precision - updated all values > References - added all

The change bars indicate the position of a technical amendment to the text with respect to the previous version of the document.

When this Instructions For Use is replaced, sign and date below and retain as specified by local regulations or laboratory policies, as appropriate.

Signature

Obsolete Date

C
Ortho-Clinical Diagnostics Mandeville House 62 The Broadway Amersham Buckinghamshire HP7 OHJ England Ortho-Clinical Diagnostics, Inc. 100 Indigo Creek Drive Rochester, NY 14626-5101

' Ortho-Clinical Diagnostics


jjjotmHm company

VITROS is a trademark of Ortho-Clinical Diagnostics, Inc. Ortho-Clinical Diagnostics, Inc., 2003. All rights reserved. Printed in USA.

Pub. No. C-307

Version 1.0

VITR
Chemistry

INSTRUCTIONS FOR USE


VITROS Chemistry Products NBIL DT Slides Intended Use

NBIL DT
Neonatal Bilirubin

For in vitro diagnostic use only. VITROS NBIL DT Slides quantitatively measure total bilirubin concentration in serum and plasma in neonates during the first weeks of life.

Summary and Explanation of the Test


Neonatal bilirubin (NBIL) is increased in erythroblastosis fetalis (hemolytic disease of the newborn), which causes jaundice in the first two days of life. Other causes of neonatal jaundice include physiologic jaundice, hematoma/hemorrhage, hypothyroidism, and obstructive jaundice.

Principles of the Procedure


The VITROS NBIL DT Slide method is performed using the VITROS NBIL DT Slide and the VITROS Chemistry Products DT Calibrator Kit on VITROS DT60/DT60 II Chemistry Systems. The VITROS NBIL DT Slide is a multilayered, analytical element coated on a polyester support. The analysis is based on a modification of the classic diazo reaction.1 A drop of patient sample is deposited on the slide and is evenly distributed by the spreading layer to the underlying layers. This layer provides a reflective background for measuring the diazo products of bilirubin and contains all reagents necessary to determine total bilirubin in neonates. The method uses dyphylline to dissociate unconjugated bilirubin from albumin. Unconjugated bilirubin and conjugated bilirubin subsequently react with the diazonium salt 4-(A/-carboxymethylsulfonyl) benzenediazonium hexafluorophosphate to produce azobilirubin chromophores that have similar molar absorptivities. The change in reflection density is proportional to the bilirubin concentration in the sample.

Reaction Sequence
neonatal bilirubin
dyphylline [4-(N-carboxymethylsulfonyl)-benzenediazonium hexafluorophosphate]

azobilirubin chromophores

Test Type and Conditions


Test Type and Conditions for NBIL DT
Test Type Colori metric VITROS DT60/DT60 II Module DT60/DT60 II Approximate Incubation Time 5 minutes Temperature 37C (98.6F) Wavelength 555 nm Sample Drop Volume 10 uL

Warnings and Precautions


For in vitro diagnostic use only. Take care when handling materials and samples of human origin. Since no test method can offer complete assurance that infectious agents are absent, consider all clinical specimens, controls, and calibrators potentially infectious. Handle specimens, solid and liquid waste, and test components in accordance with local regulations and NCCLS Guideline M292 or other published biohazard safety guidelines. For specific warnings and precautions for calibrators, quality control materials, and other components, refer to the instructions for use for the appropriate VITROS product, or to other manufacturer's product literature.

Version 1.0

Pub. No. C-364

NBILDT
Neonatal Bilirubin

INSTRUCTIONS FOR USE


Reagents

Reagents
Slide Diagram

Slide Ingredients
Reactive ingredients per cm
2
2
_ 3

Dyphylline 0.5 mg and 4-(N-carboxymethylaminosulfonyl) benzene diazonium hexafluorophosphate 57 (.ig.

Other ingredients
Pigment, binders, buffer, mordant, surfactants and stabilizer.

~~~~ ~ 4 _____

1. Upper slide mount 2. Spreading layer (BaSCXtl dyphylline diazonium salt 3. Reagent layer buffer, pH 3.0 4. Support layer * 5. Lower slide mount

Slide Handling
CMJTiGM: Do not use slides with damaged or incompletely sealed packaging.

Slide Preparation
IMPORTANT: The slide must reach room temperature, 18-28 C (64 -82 F), before the wrapper is opened. Do not use unopened slides that have been at room temperature, 18 "-28 C (64-82F) for >48 hours. 1. Remove the unopened slide from the box. 2. Warm the unopened slide for 15 minutes. Unopened slides that remain in the sealed wrapper may be returned to refrigerator or freezer storage. 3. Remove the packaging and place the slide into the loading station within 15 minutes after opening.

Slide Storage and Stability


VITROS NBIL DT Slides are stable until the expiration date on the carton when they are stored and handled as specified.

Slide Storage and Stability for NBIL DT Storage Condition Slides


Unopened Opened Room temperature Frozen Room temperature 18-28C (64-82F) <-18C (<0F) 18-28C (64-82F)

Stability <48 hours Until expiration date <15 minutes

Specimen Requirements
WARNiNG: Handle specimens as biohazardous material.

Specimens Recommended
I Serum Plasma:3 Heparin
Certain collection devices have been reported to affect other analytes and tests4 Confirm that your collection devices are compatible with this test.

Specimens Not Recommended


Samples from patients other than neonates (newborns) are not recommended. These samples should be analyzed on the VITROS TBIL DT Slide.

Serum and Plasma


Specimen Collection and Preparation
Collect specimens using standard laboratory procedures.66 Patient Preparation No special patient preparation is necessary.

Pub. No. C-364

Version 1.0

INSTRUCTIONS FOR USE


Testing Procedure

NRIL DT
Neonatal Bilirubin

Special Precautions For the effect of sample hemolysis on test results, refer to "Limitations of the Procedure." Protect specimens from light. Direct exposure to sunlight is reported to cause as much as a 50% loss of serum bilirubin in one hour, especially when the specimen is kept in capillary tubes.1 Exposure to normal laboratory light can result in a significant loss of serum bilirubin after two to three hours. Centrifuge specimens and remove the serum from the cellular material within 4 hours of collection.7

Specimen Handling and Storage


Handle specimens as biohazardous material. Handle and store specimens in stoppered containers to avoid contamination and evaporation. Mix samples by gentle inversion and bring to room temperature, 18-28C (64-82F), prior to analysis.

Specimen Storage and Stability for NBIL DT: Serum7 Temperature Storage
Room temperature Refrigerated Frozen 18-28C(64O-82F) 2-8C (36-46F) <-18C(<0F)

Stability <4 hours <7 days <6 months

Testing Procedure
Materials Provided
VITROS Chemistry Products NBIL DT Slides

Materials Required But Not Provided


VITROS Chemistry Products DT Calibrator Kit Quality control materials, such as VITROS Chemistry Products DT Control I & II VITROS Chemistry Products 7% BSA VITROS DT Pipette

Operating Instructions
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System. Bring all fluids and samples to room temperature, 18-28C (64-82F), prior to analysis. IMPORTANT:

Sample Dilution
If bilirubin concentrations exceed the system's reportable (dynamic) range: 1. Dilute the sample with VITROS 7% BSA. 2. Reanalyze. 3. Multiply the results by the dilution factor to obtain an estimate of the original sample's bilirubin concentration.

Calibration
Required Calibrators
VITROS Chemistry Products DT Calibrator Kit, bottles 1,2, and 3

Calibrator Preparation, Handling, and Storage


Refer to the instructions for use for VITROS DT Calibrator Kit.

Calibration Procedure
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

When to Calibrate
Calibrate: When the slide lot number changes. When critical system parts are replaced due to service or maintenance. When government regulations require. - For example, in the USA, CLIA regulations require calibration or calibration verification at least once every six months. Version 1.0 Pub. No. C-364

VI"TFj[Cp"S | * |

NBILDT
Neonatal Bilirubin

INSTRUCTIONS FOR USE


Quality Control

The VITROS NBIL DT test may also need to be calibrated: If quality control results are consistently outside acceptable range. After certain service procedures have been performed. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Calculations
Reflectance from the slide is measured at 555 nm after the fixed incubation time. Once a calibration has been performed for each slide lot, neonatal bilirubin concentration in unknown samples can be determined using the software-resident endpoint colorimetric math model and the response obtained from each unknown test slide.

Validity of a Calibration
Calibration parameters are automatically assessed by the VITROS DT60/DT60II Chemistry System against a set of quality parameters resident within the analyzer software. Failure to meet any of the pre-defined quality parameters results in a failed calibration. The calibration report should be used in conjunction with quality control results to determine the validity of a calibration.

Reportable (Dynamic) Range Reportable (Dynamic) Range for NBIL DT


Conventional (mg/dL) SI Units (umol/L) 2-342

0.1-20.0

For out-of-range samples, refer to "Sample Dilution."

Traceability of the Calibration


I i Values assigned to the VITROS Chemistry Products DT Calibrator Kit for neonatal bilirubin are traceable to internal Master Lots of VITROS Chemistry Products BuBc Slides and VITROS Calibrator Kit 4. Performance of the Master Lot of VITROS BuBc slides was initially established by comparison to the High Performance Liquid Chromatography (HPLC) method described by Lauff et al.8 The Ortho-Clinical Diagnostics (OCD) calibration laboratory uses the Master Lot of VITROS BuBc Slides and Master Lot of VITROS Calibrator Kit 4 for neonatal bilirubin value assignment for new lots of the VITROS DT Calibrator Kit.

Quality Control
Procedure Recommendations
| ' Handle quality control materials as biohazardous material. Choose control levels that check the clinically relevant range. Analyze quality control materials in the same manner as patient samples, before or during patient sample processing. To verify system performance, analyze control materials: - After calibration. - According to local regulations or at least once each day that the test is being performed. - After specified service procedures are performed. Refer to the operator's manual for your VITROS DT60/DT60 II System. If control results fall outside your acceptable range, investigate the cause before deciding whether to report patient results. For general quality control recommendations, refer to Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition9 or other published guidelines. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Quality Control Material Selection


| | IMPORTANT: VITROS DT Control I & II are recommended for use with the VITROS DT60/DT60 II Chemistry System. Evaluate the performance of other commercial control fluids for compatibility with this test before using for quality control.

Control materials other than VITROS DT Controls I & II may show a difference when compared with other neonatal bilirubin methods if they: - Depart from a true human matrix. - Contain high concentrations of preservatives, stabilizers, or other nonphysiological additives. Do not use control materials stabilized with ethylene glycol.

Pub. No. C-364

Version 1.0

INSTRUCTIONS FOR USE


Expected Values and Reporting Units

NBIIDT
Neonatal Bilirubin

Quality Control Material Preparation and Storage


Refer to the instructions for use for VITROS Chemistry Products DT Control I & II or to other manufacturer's product literature.

Expected Values and Reporting Units


Reference Interval
This reference interval is the central 95% of results from a study of 40 apparently healthy neonates (average age 1.2 days) with normal liver enzymes.

Reference Interval for NBIL DT


Conventional Units (mg/dL) Neonates SI Units (umol/L) 17-180

1.0-10.5

Each laboratory should confirm the validity of these intervals for the population it serves.

Reporting Units and Unit Conversion


The VITROS DT60/DT60 II Chemistry System may be programmed to report neonatal bilirubin results in conventional and SI units.

Reporting Units and Unit Conversion for NBIL DT Conventional Units SI Units
mg/dL umol/L (mg/dL x 17.1)

Limitations of the Procedure


Known Interferences
The VITROS NBIL Slide method was screened for interfering substances following NCCLS Protocol EP7.10 The substances listed in the table, when tested at the concentrations indicated, caused the bias shown.

Known Interfering Substances for NBIL DT


Interferent* Levodopa 4-Aminosalicylic acid Phenazopyridine Biliverdin Hemoglobin Interferent Concentration 300 ug/mL** 300 |.ig/mL** 8 mg/dL 8 mg/dL 8 mg/dL 8 mg/dL 4 mg/dL 4 mg/dL 100 mg/dL 200 mg/dL 400 mg/dL 100 mg/dL 200 mg/dL 400 mg/dL 100 mg/dL 200 mg/dL 400 mg/dL Bilirubin Cone. SI (Mmol/L) Conv. (mg/dL) 0.4 (1.52mmol/L)** 7 (1.52 mmol/L)** 17.8 304 (0.52 mmol/L) 0.4 7 (0.52 mmol/L) 17.6 301 (0.3 mmol/L) 0.4 7 (0.3 mmol/L) 17.4 298 0.3 (69 umol/L) 5 16.4 (69 umol/L) 280 0.5 9 d g/L) 0.5 9 (2 g/L) 0.5 9 (4 g/L) 10.5 180 (1 g/L) 10.5 (2 g/L) 180 10.5 (4 g/L) 180 15.0 257 (1 g/L) 15.0 257 (2 g/L) 15.0 (4 g/L) 257 Bias SI Conv. (mg/dL) +1.4 -9.4 +0.4 +2.5 +3.8 +3.5 +0.5 +0.7 +0.1 +0.4 + 1.3 -1.0 -1.2 -1.0 -1.2 -1.7 -1.9 (Mmol/L) +24 -161 +7 +43 +65 +60 +9 +12 +2 +7
+22

-17 -21 -17 -21 -29 -32

It is possible that other interfering substances may be encountered. These results are representative; however, your results may differ somewhat due to test-to-test variation. The degree of interference at concentrations other than those listed might not be predictable. This concentration may be present in the treatment of Parkinson's disease.

Version 1.0

Pub. No. C-364

NBIL DT
Neonatal Bilirubin

INSTRUCTIONS FOR USE


Performance Characteristics It is important to maintain consistency in bilirubin methodology. If VITROS NBIL DT Slides are initially used to monitor a patient, continue to monitor that patient with VITROS NBIL DT Slides. Do not switch to VITROS TBIL DT Slides regardless of the patient's age. Results from the VITROS NBIL DT Slide may not be accurate at elevation greater than 6000 feet (approx. 1800 meters) above sea level. Cefotiam (Pansporin) has been reported to show very large positive biases on VITROS NBIL DT Slide results.11 This drug is normally cleared through the kidney. Biases will be largest in specimens from patients with renal insufficiency and may be as large as 5 mg/dL (86 |.imol/L). Drugs and other compounds that are diazo-reactive or that absorb light in the vicinity of 555 nm may interfere. Certain drugs and clinical conditions are known to alter total bilirubin concentration in vivo. For additional information, refer to one of the published summaries.1213

Other Limitations

Performance Characteristics
Method Comparison
I I The plot and table show the results of a comparison of samples analyzed on the VITROS DT60 II System with those analyzed using the VITROS 950 System and VITROS BuBc Slides to calculate NBIL. Testing followed NCCLS Protocol EP9.14 Method Comparison for NBIL DT: Serum Conventional Units 25 400
SI Units

t
E

20
!* >

300

200 f
Q W

10
O a: >

100

10

15

20

100

200

300

400

Comparative Method: VITROS 950 System (mg/dL)

Comparative Method: VITROS 950 System (Mmol/L)

Method Comparison for NBIL DT: Serum


Conventional Units (mg/dL) n DT60 II System vs. 950 System 104 Correlation Slope Coefficient 0.99 0.998 Range of Sample Cone. 0.1-18.2 Intercept 0.10 Sy.x 0.32 SI Units (umol/L) Range of Sample Cone. 2-312 Intercept 1.63 Sy.x 5.48

Precision
Precision was evaluated with quality control materials on the VITROS DT60 II System following NCCLS Protocol EP5.15 The data presented are a representation of test performance and are provided as a guideline. Variables such as sample handling and storage, reagent handling and storage, laboratory environment, and system maintenance can affect reproducibility of test results. Precision for NBIL DT: Serum Conventional Units (mg/dL) Within Within Mean Cone. Day SD* Lab SD** 1.3 14.5 0.10 0.22 0.10 0.29 SI Units (Mmol/L) Mean Cone. 23 248 Within Day SD* 1.6 3.8 Within Lab SD** 1.8 5.0 Within Lab CV%** 7.8 2.0 No. Observ. 84 84 No. Days 21 21

System VITROS DT60 II

Within Day precision was determined using two runs/day with two replications. Within Lab precision was determined using a single lot of slides and calibrating weekly.

Pub.

No. C-364

Version 1.0

INSTRUCTIONS FOR USE


References

NBILDT
Neonatal Bilirubin

References
1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. Routh Jl: Liver Function. In Tietz NW(ed). Fundamentals of Clinical Chemistry, ed 2. Philadelphia; WB Saunders; 1037; 1976. NCCLS. Protection of Laboratory Workers from Instrument Biohazards and Infectious Diseases Transmitted by Blood, Body Fluids and Tissue: Approved Guideline. NCCLS Document M29 (OSBN 1-56238). NCCLS, Wayne, PA 19087; 1997. Doumas BT, et al. Differences Between Values for Plasma and Serum in Tests Performed in the Ektachem 700 XR Analyzer, and Evaluation of "Plasma Separator Tubes (PST)." Clin. Chem. 35:151-153; 1989. Calam RR. Specimen Processing Separator Gels: An Update. J Clin Immunoassay. 11:86-90; 1988. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Venipuncture. NCCLS Document H3. Wayne, PA: NCCLS; 1991. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Skin Puncture. NCCLS Document H4. Wayne, PA: NCCLS; 1991. Clinical Laboratory Handbook for Patient Preparation and Specimen Handling. Fascicle VI: Chemistry/Clinical Microscopy. Northfield, IL: College of American Pathologists; 1992. Lauff JJ, Kasper ME, Wu T.W, Ambrose RT. Isolation and preliminary characterization of a fraction of bilirubin in serum that is firmly bound to protein. Clin. Chem. 28:629-637, 1982. NCCLS. Statistical Quality Control for Quantitative Measurements: Principles and Definitions: Approved Guideline-Second Edition. NCCLS Document C24. Wayne, PA: NCCLS; 1999. NCCLS. Interference Testing in Clinical Chemistry. NCCLS Document EP7. Wayne, PA: NCCLS; 1986. Pickert A, Riedlinger I, Stumvill M. Interference of Cefotiam with Total Bilirubin Measured with the Ektachem Analyzer. Clin. Chem. 38:599-600; 1992. Young DS. Effects of Drugs on Clinical Laboratory Tests, ed. 4. Washington D.C.: AACC Press; 1995. Friedman RB, Young DS. Effects of Disease on Clinical Laboratory Tests. Washington, D.C.: AACC Press; 1990. NCCLS. Method Comparison and Bias Estimation Using Patient Samples: Approved Guideline. NCCLS Document EP9. Wayne, PA: NCCLS; 1995. NCCLS. User Evaluation of Precision Performance with Clinical Chemistry Devices. NCCLS Document EP5. Wayne, PA: NCCLS; 1992.

Glossary of Symbols
The Glossary of Symbols table defines the symbols used on the VITROS Chemistry Products packaging and on the VITROS Chemistry Products Assay Sheets.

Glossary of Symbols
Do Not Reuse Use by or Expiration Date (YYYY-MM-DD) Lot Number BC mf I Manufacturer Authorized Representative Store Between Consult Instructions for Use Fragile, Handle with Care. Keep Dry This end up

\ y /

Tests

Contains Sufficient for "n"

Q|L| On!
"^rzi MfcX [

Manufacturer's Serial Number


Catalog Number or Product Code Attention: See Instructions for Use.

For In Vitro Diagnostic Use

Store At or Below Store At or Above

Version 1.0

Pub. No. C-364

NBILDT
Neonatal Bilirubin

INSTRUCTIONS FOR USE


Revision History

Revision History
Date of Revision 2003-10-01 Version 1.0 Description of Technical Changes* > New format > New organization and sections consistent with IVD Directive > Known Interfering Substances - updated values; added phenazopyridine and hemoglobin; changed 5-aminosalicylic acid to 4-aminosalicylic acid > Method Comparison - updated all comparisons and the plots > Precision - updated all values > References - added all except 6, 12,13

The change bars indicate the position of a technical amendment to the text with respect to the previous version of the document.

When this Instructions For Use is replaced, sign and date below and retain as specified by local regulations or laboratory policies, as appropriate.

Signature

Obsolete Date

C
I EC j REP I
Ortho-Clinical Diagnostics Mandeville House 62 The Broadway Amersham Buckinghamshire HP7 OHJ England Ortho-Clinical Diagnostics, Inc. 100 Indigo Creek Drive Rochester, NY 14626-5101

'Ortho-Clinical Diagnostics
company

VITROS is a trademark of Ortho-Clinical Diagnostics, Inc. Ortho-Clinical Diagnostics, Inc., 2003. All rights reserved. Printed in USA.

Pub. No. C-364

Version 1.0

VITR
Chemhtry

INSTRUCTIONS FOR USE


VITROS Chemistry Products NH3 DT Slides Intended Use
For in vitro diagnostic use only. VITROS NH3 DT Slides quantitatively measure ammonia (NH3) concentration in plasma.

NHsDT
Ammonia

Summary and Explanation of the Test


Ammonia is a waste product of protein catabolism; it is potentially toxic to the central nervous system. Increased plasma ammonia may be indicative of hepatic encephalopathy, hepatic coma in terminal stages of liver cirrhosis, hepatic failure, acute and subacute liver necrosis, and Reye's syndrome. Hyperammonemia may also be found with increasing dietary protein intake.1 In addition to its use for endogenous ammonia determination, the VITROS NH3 DT Slides are also used in conjunction with VITROS CREA DT Slides for creatinine as described in the VITROS Chemistry Products CREA DT Instructions for Use (blankcorrected method).

Principles of the Procedure


The VITROS NH3 DT Slide method is performed using the VITROS NH3 DT Slide and the VITROS Chemistry Products DT Calibrator Kit on VITROS DT60/DT60 II Chemistry Systems. The VITROS NH3 DT Slide is a multilayered, analytical element coated on a polyester support. A drop of patient sample is deposited on the slide and is evenly distributed by the spreading layer to the underlying layers. Water and nonproteinaceous components travel to the underlying buffered reagent layer, and the ammonium ions are converted to gaseous ammonia. The semipermeable membrane allows only ammonia to pass through and prevents buffer or hydroxyl ions from reaching the indicator layer. After a fixed incubation period, the reflection density of the dye is measured using the white background of the spreading layer as a diffuse reflector.

Reaction Sequence
NH3 + bromphenol blue (ammonia indicator) blue dye

Test Type and Conditions


Test Type and Conditions for NH3 DT
Test Type Colorimetric VITROS DT60/DT60 II Module DT60/DT60 II Approximate Incubation Time 5 minutes Temperature 37C (98.6F) Wavelength 605 nm Sample Drop Volume 10 uL

Warnings and Precautions


For in vitro diagnostic use only. Take care when handling materials and samples of human origin. Since no test method can offer complete assurance that infectious agents are absent, consider all clinical specimens, controls, and calibrators potentially infectious. Handle specimens, solid and liquid waste, and test components in accordance with local regulations and NCCLS Guideline M292 or other published biohazard safety guidelines. For specific warnings and precautions for calibrators, quality control materials, and other components, refer to the instructions for use for the appropriate VITROS product, or to other manufacturer's product literature.

Version 1.0

Pub. No. C-333

NHaDT
Ammonia

INSTRUCTIONS FOR USE


Reagents

Reagents
Slide Diagram

Slide Ingredients Reactive ingredients per cm


Bromphenol blue 27 pg.
2

Other ingredients
Pigment, binders, surfactants, buffer and stabilizer.
"-

-4 5

1. Upper slide mount 2. Spreading layer (TIO2) 3. Reagent layer buffer, pH 9.3 4. Semlpermeable membrane 5. Indicator layer bromphenol blue 6. Support layer 7. Lower slide mount

Slide Handling
N: Do not use slides with damaged or incompletely sealed packaging.

Slide Preparation
The slide must reach room temperature, 18-28 C (64 -82 F), before the wrapper is opened. Do not use unopened slides that have been at room temperature, 18 - 2 8 C (64 -82 F) for >48 hours. 1. 2. Remove the unopened slide from the box. Warm the unopened slide for 15 minutes. Unopened slides that remain in the sealed wrapper may be returned to refrigerator or freezer storage. 3. Remove the packaging and place the slide into the loading station within 15 minutes after opening.

Slide Storage and Stability


VITROS NH? DT Slides are stable until the expiration date on the carton when they are stored and handled as specified.

Slide Storage and Stability for NH3 DT Storage Condition Slides


Unopened Room temperature Refrigerated Frozen Room temperature 18-28C (64-82F) 2-8C (36-46F) <-18C (<0F) 18-28C (64-82F)

Opened

Stability <48 hours <4 weeks Until expiration date <15 minutes

Specimen Requirements
WARNING: Handle specimens as biohazardous material.

Specimens Recommended for Measurement of Ammonia


I I Plasma:3 EDTA Meparin (except ammonia heparin) Serum should not be used for ammonia measurements because ammonia is produced during the clotting process.3 Certain collection devices have been reported to affect other analytes and tests." Confirm that your collection devices are compatible with this test.

NOTE:

I I

IMPORTANT:

Specimens Recommended for Measurement of Creatinine Where Ammonia Serves as Blank


I | Plasma:3 Serum Certain collection devices have been reported to affect other analytes and tests4 Confirm that your collection devices are compatible with this test. EDTA Heparin (except ammonia heparin)

IMPORTANT:

Pub. No. C-333

Version 1.0

INSTRUCTIONS FOR USE


Testing Procedure Specimens Not Recommended Do not use hemolyzed specimens.3 Plasma or Serum Specimen Collection and Preparation
Piasma Collect specimens using standard laboratory procedures.5 6 Keep on ice until analysis.5 6 Serum Collect specimens using standard laboratory procedures.5 6 Do not put specimens on ice. Patient Preparation No special patient preparation is necessary.

NHsDT
Ammonia

Special Precautions NOTE: Avoid using ammonia-containing cleaning solutions or hand creams in the area around the analyzer. Centrifuge specimens and remove the plasma from the cellular material within 15 minutes of collection.3 Handling and Storage Handle specimens as biohazardous material.

Specimen \fi

Handle and store specimens in stoppered containers to avoid contamination and evaporation. Mix samples by gentle inversion and bring to room temperature, 18-28C (64-82F); analyze immediately.

Specimen Storage and Stability for NH3 DT: Plasma3 Storage Temperature
Room temperature Refrigerated Frozen 18-28C(64O-82F) 2-8C (36-46F) <-18C(<0F)

Stability not recommended <3 hours <24 hours

Testing Procedure
Materials Provided
. VITROS Chemistry Products NH3 DT Slides

Materials Required But Not Provided


VITROS Chemistry Products DT Calibrator Kit Quality control materials, such as VITROS Chemistry Products DT Control I & II Isotonic saline Reagent-grade water VITROS DT Pipette

Operating Instructions
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System. Bring all fluids and samples to room temperature, 18-28C (64-82F); analyze immediately. IMPORTANT:

Sample Dilution
If ammonia concentrations exceed the system's reportable (dynamic) range: 1. Dilute the sample with isotonic saline or reagent-grade water. 2. Reanalyze. 3. Multiply the results by the dilution factor to obtain an estimate of the original sample's ammonia concentration.

Version 1.0

Pub. No. C-333

NH.DT
Ammonia

INSTRUCTIONS FOR USE


Calibration

Calibration
Required Calibrators
VITROS Chemistry Products DT Calibrator Kit, bottles 1, 2, 3, and 4

Calibrator Preparation, Handling, and Storage


NOTE: To avoid ammonia formation, calibrator fluids should be prepared only when ready to calibrate and should be run within 1 hour after preparation. Because ammonia is produced in the VITROS BUN/UREA DT Slide reaction, the analyzer will display a warning message "ANALYER READYNO CREA/NH3" during calibration of VITROS BUN/UREA DT Slides. Therefore, it is recommended that creatinine and ammonia be the first tests calibrated after the preparation of calibrator fluids.

NOTE;

Refer to the instructions for use for VITROS DT Calibrator Kit.

Calibration Procedure
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

When to Calibrate
NOTE: The VITROS CREA DT test is dependent on correct calibration of the VITROS NH3 DT Slides used as blanks. Therefore, the NH3 DT Slides must be calibrated whenever CREA DT Slides are calibrated.

Calibrate: When the slide lot number changes. When critical system parts are replaced due to service or maintenance. When government regulations require. - For example, in the USA, CLIA regulations require calibration or calibration verification at least once every six months. The VITROS NH3 DT test may also need to be calibrated: If quality control results are consistently outside acceptable range. After certain service procedures have been performed. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Calculations

Reflectance from the slide is measured at 605 nm after the fixed incubation time. Once a calibration has been performed for each slide lot, ammonia concentration in unknown samples can be determined using the software-resident endpoint colorimetric math model and the response obtained from each unknown test slide.

Validity of a Calibration

Calibration parameters are automatically assessed by the VITROS DT60/DT60 II Chemistry System against a set of quality

parameters resident within the analyzer software. Failure to meet any of the pre-defined quality parameters results in a failed calibration. The calibration report should be used in conjunction with quality control results to determine the validity of a calibration.

Reportable (Dynamic) Range Reportable (Dynamic) and SI Units NH3 DT Conventional Range for (umol/L) 1-500
For out-of-range samples, refer to "Sample Dilution."

Traceability of the Calibration


Values assigned to the VITROS Chemistry Products DT Calibrator Kit for ammonia are traceable to gravimetrically prepared standards made from reagent grade ammonium sulfate. The Ortho-Clinical Diagnostics calibration laboratory uses the gravimetrically prepared standard to calibrate an enzymatic procedure using glutamate dehydrogenase to measure ammonia7 to support value assignment for the VITROS DT Calibrator Kit.

Pub. No. C-333

Version 1.0

INSTRUCTIONS FOR USE


Quality Control

NHsDT
Ammonia

Quality Control
Procedure Recommendations
| : llfJG Handle quality control materials as biohazardous material. Choose control levels that check the clinically relevant range. Analyze quality control materials in the same manner as patient samples, before or during patient sample processing. To verify system performance, analyze control materials: - After calibration. - According to local regulations or at least once each day that the test is being performed. - After specified service procedures are performed. Refer to the operator's manual for your VITROS DT60/DT60 II System. If control results fall outside your acceptable range, investigate the cause before deciding whether to report patient results. For general quality control recommendations, refer to Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition8 or other published guidelines. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Quality Control Material Selection

I
I

IMPORTANT;

VITROS DT Control I & II are recommended for use with the VITROS DT60/DT60 II

Chemistry System. Evaluate the performance of other commercial control fluids for compatibility with this test before using for quality control. Control materials other than VITROS DT Controls I & II may show a difference when compared with other ammonia methods if they: - Depart from a true human matrix. - Contain high concentrations of preservatives, stabilizers, or other nonphysiological additives. Do not use control materials stabilized with ethylene glycol.

Quality Control Material Preparation and Storage


Refer to the instructions for use for VITROS Chemistry Products DT Control I & II or to other manufacturer's product literature.

Expected Values and Reporting Units


Reference Interval
This reference interval is the central 95% of results from an internal study of 60 apparently healthy adults from a working population (34 females and 26 males). No significant differences between results from the male and female populations were observed.

Reference Interval for NH3 OT


Conventional and SI Units (umol/L)

9-30
Each laboratory should confirm the validity of these intervals for the population it serves.

Reporting Units and Unit Conversion


The VITROS DT60/DT60 II Chemistry System may be programmed to report ammonia results in conventional and SI units. Reporting Units for NH 3 DT Conventional and SI Units umol/L

Version 1.0

Pub. No. C-333

NHaDT
Ammonia

INSTRUCTIONS FOR USE


Limitations of the Procedure

Limitations of the Procedure


Known Interferences
I Glucose at 600 mg/dL (33.3 mmol/L) may cause a decrease of 40 umol/L j n ammonia concentration. If an NH3 slide follows a BUN/UREA DT Slide immediately, high BUN values may increase ammonia values. - A BUN value up to 40 mg/dL (14.3 mmol/L) may increase ammonia values by 34 umol/L. - BUN values over 40 mg/dL (14.3 mmol/L) will cause the analyzer to print an R18 code next to the ammonia result. Discard the result and repeat the sample without the BUN/UREA DT Slide in the incubator. NH3 DT testing should not be done when any BUN/UREA DT Slides are in the incubator.

IMPORTANT:

Other Limitations
Certain drugs and clinical conditions are known to alter ammonia concentration in vivo. For additional information, refer to one of the published summaries.9'1C

Performance Characteristics
Method Comparison

The plot and table show the results of a comparison of samples analyzed on the VITROS DT60 II System with those analyzed using the VITROS 950 System. Testing followed NCCLS Protocol EP9.11

Method Comparison for NH3 DT


Conventional and SI Units 500 400

300 200

o
oc I00

100

200

300

4li(i

500

Comparative Method: VITROS 950 System (Mmol/L)

Method Comparison for NH3 DT


Conventional and SI Units (Mmol/L) n DT60 II System vs. 950 System 59 Slope 1.02 Correlation Coefficient 0.997 Range of Sample Cone. Intercept 3.71 Sy.x 10.73

5-460

Pub. No. C-333

Version 1.0

gj VITRCpS

INSTRUCTIONS FOR USE


References

NHaDT
Ammonia

Precision
Precision was evaluated with quality control materials on the VITROS DT60 II System following NCCLS Protocol EP5.12 The data presented are a representation of test performance and are provided as a guideline. Variables such as sample handling and storage, reagent handling and storage, laboratory environment, and system maintenance can affect reproducibility of test results.

Precision for NH3DT


Conventional and SI Units (pmol/L) System VITROS DT60 II Mean Cone. 69 235 Within Day SD* 6.3 14.6 Within Lab SD** 6.8 16.2 Within Lab CV%** 10.0 6.9 No. Observ. 88 88 No. Days 22 22

Within Day precision was determined using two runs/day with two replications. Within Lab precision was determined using a single lot of slides and calibrating weekly.

References
Tietz NW(ed). Fundamentals of Clinical Chemistry, ed. 3. Philadelphia: WB Saunders; 748; 1987. NCCLS. Protection of Laboratory Workers from Instrument Biohazards and Infectious Diseases Transmitted by Blood, Body Fluids and Tissue; Approved Guideline. NCCLS Document M29 (OSBN 1-56238). NCCLS, Wayne, PA 19087; 1997. 3. Clinical Laboratory Handbook for Patient Preparation and Specimen Handling. Fascicle VI: Chemistry/Clinical Microscopy. Northfield, IL: College of American Pathologists; 1992. 4. Calam RR. Specimen Processing Separator Gels: An Update. J Clin Immunoassay. 11:86-90; 1988. 5 NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Venipuncture. NCCLS Document H3. Wayne, PA: NCCLS; 1991. 6. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Skin Puncture. NCCLS Document H4. Wayne, PA: NCCLS; 1991. 7. Bruce WA, Leiendecker, CM, Freier EF. Two-Point Determination of Plasma Ammonia with the Centrifugal Analyzer. Clin. Chem. 24:782; 1978. 8. NCCLS. Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition. NCCLS Document C24. Wayne, PA: NCCLS; 1999. 9. Young DS. Effects of Drugs on Clinical Laboratory Tests, ed. 4. Washington D.C.: AACC Press; 1995. 10. Friedman RB, Young DS. Effects of Disease on Clinical Laboratory Tests. Washington, D.C.: AACC Press; 1990. 11. NCCLS. Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline. NCCLS Document EP9. Wayne, PA: NCCLS; 1995. 12. NCCLS. User Evaluation of Precision Performance with Clinical Chemistry Devices. NCCLS Document EP5. Wayne, PA: NCCLS; 1992. 1. 2.

Glossary of Symbols
The Glossary of Symbols table defines the symbols used on the VITROS Chemistry Products packaging and on the VITROS Chemistry Products Assay Sheets.

Glossary of Symbols
Do Not Reuse Use by or Expiration Date (YYYY-MM-DD) Lot Number Manufacturer's Serial Number Catalog Number or Product Code Attention: See Instructions for Use. [ EC [ na> [ \ v/ V Manufacturer Authorized Representative Contains Sufficient for "n" Tests For In Vitro Diagnostic Use Store At or Below Store At or Above Store Between Consult Instructions for Use

Fragile, Handle with Care. Keep Dry

It

This end up

Version 1.0

Pub. No. C-333

NHsDT
Ammonia

INSTRUCTIONS FOR USE


Revision History

Revision History
Date of Revision 2003-10-01 Version 1.0 Description of Technical Changes* > New format > New organization and sections consistent with IVD Directive > Specimens Recommended - updated wording for heparin; added EDTA Specimen Storage and Stability - updated stability values > Reference Interval - updated upper limit > Limitations of the Procedure - updated interferent statements > Method Comparison - updated all comparisons and the plot > Precision - updated all data > References - added all

The change bars indicate the position of a technical amendment to the text with respect to the previous version of the document. When this Instructions For Use is replaced, sign and date below and retain as specified by local regulations or laboratory policies, as appropriate.

Signature

Obsolete Date

C
EC i REP i

Ortho-Clinical Diagnostics Mandeville House 62 The Broadway Amersham Buckinghamshire HP7 OHJ England Ortho-Clinical Diagnostics, Inc. 100 Indigo Creek Drive Rochester, NY 14626-5101

' Ortho-Clinical Diagnostics


a ofMttm^o&Mfm company

VITROS is a trademark of Ortho-Clinical Diagnostics, Inc. Ortho-Clinical Diagnostics, Inc., 2003. All rights reserved. Printed in USA.

Pub. No. C-333

Version 1.0

Products

VITR
Chemistry

INSTRUCTIONS FOR USE


VITROS Chemistry Products PHOS DT Slides Intended Use

PHOS DT
Phosphorus

For in vitro diagnostic use only. VITROS PHOS DT Slides quantitatively measure phosphorus (PHOS) concentration in serum and plasma.

Summary and Explanation of the Test


Phosphorus, as phosphate, is distributed throughout the body. Causes of high serum phosphorus include dehydration, hypoparathyroidism, hypervitaminosis D, metastases to bone, sarcoidosis, pulmonary embolism, renal failure, and diabetes mellitus with ketosis. Low serum phosphorus is found in primary hyperparathyroidism and other causes of serum calcium elevation, sepsis, vitamin D deficiency, renal tubular disorders, chronic hemodialysis, vomiting, and occasionally with decreased dietary phosphate intake.1

Principles of the Procedure


The VITROS PHOS DT Slide method is performed using the VITROS PHOS DT Slide and the VITROS Chemistry Products DT Calibrator Kit on VITROS DT60/DT60 II Chemistry Systems. The VITROS PHOS DT Slide is a muitiiayered, analytical element coated on a polyester support. The analysis is based on the reaction of inorganic phosphate with ammonium molybdate to form an ammonium phosphomolybdate complex at acidic pH, as described by Fiske and Subbarow.2 p-Methylaminophenol sulfate, an organic reductant reported by Gomori,3 reduces the complex to form a stable heteropoiymolybdenum blue chromophore. A drop of patient sample is deposited on the slide and is evenly distributed by the. spreading layer to the underlying layers. Phosphorus in the specimen forms a complex with ammonium moiybdate. This complex is reduced by p-methylaminophenol sulfate to give a blue complex. The concentration of phosphorus in the sample is determined by measuring the heteropoiymolybdenum blue complex by reflectance spectrophotometry.

Reaction Sequence
inorganic phosphate + ammonium molybdate ammonium phosphomolybdate complex
pH4.2

> ammonium phosphomolybdate complex -> heteropolymolybdate blue

p-methylaminophenol sulfate

Test Type and Conditions


Test Type and Conditions for PHOS DT
Test Type Colorimetric VITROS DT60/DT60 II Module DT60/DT60 II Approximate Incubation Time 5 minutes Temperature 37C (98.6F) Wavelength 660 nm Sample Drop Volume 10 ML

Warnings and Precautions


For in vitro diagnostic use only. Take care when handling materials and samples of human origin. Since no test method can offer complete assurance that infectious agents are absent, consider all clinical specimens, controls, and calibrators potentially infectious. Handle specimens, solid and liquid waste, and test components in accordance with local regulations and NCCLS Guideline M294 or other published biohazard safety guidelines. For specific warnings and precautions for calibrators, quality control materials, and other components, refer to the instructions for use for the appropriate VITROS product, or to other manufacturer's product literature.

Version 1.0

Pub. No. C-350

PHOS DT
Phosphorus

INSTRUCTIONS FOR USE


Reagents

Reagents
Slide Diagram

Slide Ingredients
Reactive ingredients per cm

I 2

p-methylaminophenol sulfate 350 ug; and ammonium molybdate 340 ug.

Other ingredients
Pigment, binders, surfactants, buffer and stabilizers.
-__

^__
s

1. Upper slide mount 2. Spreading layer (BaSO4) 3. Reagent layer p-methylaminophenol sulfale ammonium molybdate buffer, pH 4.2 4. Support layer 6. Lower slide mount

.. - -

Slide Handling
CAUTiOH: Do not use slides with damaged or incompletely sealed packaging.

Slide Preparation
IMPORTANT: The slide must reach room temperature, 18 -28 C (64 "-82 F), before the wrapper is opened. Do not use unopened slides that have been at room temperature, 18 -28 "C (64-82F) for >48 hours. 1. 2. Remove the unopened slide from the box. Warm the unopened slide for 15 minutes. Unopened slides that remain in the sealed wrapper may be returned to refrigerator or freezer storage. 3. Remove the packaging and place the slide into the loading station within 15 minutes after opening.

Slide Storage and Stability


VITROS PHOS DT Slides are stable until the expiration date on the carton when they are stored and handled as specified.

Slide Storage and Stability for PHOS DT


Slides Unopened Storage Condition Room temperature 18-28C(64-82 F) Refrigerated 2-8C (36-46F) Frozen <-18C(<0F) 18-28C(64-82 F) Room temperature Stability <48 hours Until expiration date Until expiration date <15 minutes

Opened

Specimen Requirements
WARNING: Handle specimens as biohazardous material.

Specimens Recommended
Serum Plasma:5 Heparin Certain collection devices have been reported to affect other analytes and tests.e Confirm that your collection devices are compatible with this test.

IMPORTANT:

Specimens Not Recommended


. EDTA Fluoride oxalate Citrate Do not use hemolyzed specimens. Hemolysis produces elevated phosphorus values due to the inorganic phosphates and phosphatases present in red blood cells.7 Plasma:7

Pub.

No. C-350

Version 1.0

gl VITRI

INSTRUCTIONS FOR USE


Testing Procedure Serum and Plasma Specimen Collection and Preparation
Collect specimens using standard laboratory procedures.8 9 Patient Preparation No special patient preparation is necessary. Special Precautions Centrifuge specimens and remove the serum from the cellular material within 4 hours of collection.10 Specimen Handling and Storage Handle specimens as biohazardous material. Handle and store specimens in stoppered containers to avoid contamination and evaporation. Mix samples by gentle inversion and bring to room temperature, 18-28C (64-82F), prior to analysis. Specimen Storage and Stability for PHOS PT: Serum and Plasma10 Storage Room temperature Refrigerated Frozen Temperature 18-28 o C(64-82 0 F) 2-8C (36-46F) <-18C (<0F) Stability <3 days <7 days <2 months

PHOS DT
Phosphorus

Testing Procedure
Materials Provided
VITROS Chemistry Products PHOS DT Slides

Materials Required But Not Provided


. VITROS Chemistry Products DT Calibrator Kit Quality control materials, such as VITROS Chemistry Products DT Control I & II Isotonic saline Reagent-grade water VITROS DT Pipette

Operating Instructions
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System. Bring all fluids and samples to room temperature, 18-28C (64-82F), prior to analysis. IMPORTANT:

Sample Dilution
If phosphorus concentrations exceed the system's reportable (dynamic) range: 1. Dilute with isotonic saline or reagent-grade water. 2. Reanalyze. 3. Multiply the results by the dilution factor to obtain an estimate of the original sample's phosphorus concentration.

Calibration
Required Calibrators .
VITROS Chemistry Products DT Calibrator Kit, bottles 1, 2, and 4

Calibrator Preparation, Handling, and Storage


Refer to the instructions for use for VITROS DT Calibrator Kit.

Calibration Procedure
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Version 1.0

Pub. No. C-350

PHOSDT
Phosphorus

INSTRUCTIONS FOR USE


Quality Control

When to Calibrate
Calibrate: When the slide lot number changes. When critical system parts are replaced due to service or maintenance. When government regulations require. - For example, in the USA, CLIA regulations require calibration or calibration verification at least once every six months. The VITROS PHOS DT test may also need to be calibrated: If quality control results are consistently outside acceptable range. After certain service procedures have been performed. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Calculations
Reflectance from the slide is measured at 660 nm after the fixed incubation time. Once a calibration has been performed for each slide lot, phophorous concentration in unknown samples can be determined using the software-resident endpoint colorimetric math model and the response obtained from each unknown test slide.

Validity of a Calibration
Calibration parameters are automatically assessed by the VITROS DT60/DT60 II Chemistry System against a set of quality parameters resident within the analyzer software. Failure to meet any of the pre-defined quality parameters results in a failed calibration. The calibration report should be used in conjunction with quality control results to determine the validity of a calibration.

Reportable (Dynamic) Range Reportable (Dynamic) Range for PHOS DT


SI Units (mmol/L) 0.16-4.20 For out-of-range samples, refer to "Sample Dilution." Conventional (mg/dL)

0.5-13.0

Traceability of the Calibration


Values assigned to the VITROS Chemistry Products DT Calibrator Kit for phosphorus are traceable to the Certified NIST (National Institute of Standards and Technology) Reference Material, SRM (Standard Reference Material) 200. The Ortho-Clinical Diagnostics (OCD) calibration laboratory uses SRM200 to calibrate the phosphomolybdate/phenylenediamine method11 to support value assignment for the VITROS DT Calibrator Kit.

Quality Control
Procedure Recommendations
| " Handle quality control materials as biohazardous material. Choose control levels that check the clinically relevant range. Analyze quality control materials in the same manner as patient samples, before or during patient sample processing. To verify system performance, analyze control materials: - After calibration. - According to local regulations or at least once each day that the test is being performed. - After specified service procedures are performed. Refer to the operator's manual for your VITROS DT60/DT60 II System. If control results fall outside your acceptable range, investigate the cause before deciding whether to report patient results. For general quality control recommendations, refer to Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition12 or other published guidelines. For additional information, refer to the operator's manual for your VITROS DT60/DT60II Chemistry System.

Pub. No. C-350

Version 1.0

INSTRUCTIONS FOR USE


Expected Values and Reporting Units

PHOS DT
Phosphorus

Quality Control Material Selection


IMPORTANT: VITROS DT Control I & II are recommended for use with the VITROS DT60/DT60 II Chemistry System. Evaluate the performance of other commercial control fluids for compatibility with this test before using for quality control.

Control materials other than VITROS DT Controls I & II may show a difference when compared with other phosphorus methods if they: - Depart from a true human matrix. - Contain high concentrations of preservatives, stabilfzers, or other nonphysiological additives. Do not use control materials stabilized with ethylene glycol.

Quality Control Material Preparation and Storage


Refer to the instructions for use for VITROS Chemistry Products DT Control I & II or to other manufacturer's product literature.

Expected Values and Reporting Units


Reference Interval
The reference interval is the central 95% of results from an internal study of 297 apparently healthy adults from a working population.

Reference Interval for PHOS DT Conventional Units (mg/dL) 2.5-4.5

SI Units (mmol/L) 0.81-1.45

Each laboratory should confirm the validity of these intervals for the population it serves.

Reporting Units and Unit Conversion


The VITROS DT60/DT60 II Chemistry System may be programmed to report phosphorus results in conventional and SI units.

Reporting Units and Unit Conversion for PHOS DT Conventional Units SI Units
mg/dL mmol/L (mg/dL x 0.3229)

Limitations of the Procedure


Known Interferences
Mannitol may increase phosphorus results. At 5 mg/dL (1.6 mmol/L) phosphorus, 640 mg/dL (35 mmol/L) D-mannitol caused a 10% increase.

Other Limitations
Certain drugs and clinical conditions are known to alter phosphorus concentration in vivo. For additional information, refer to one of the published summaries.13 u

Version 1.0

Pub. No. C-350

PHOSDT
Phosphorus

INSTRUCTIONS FOR USE


Performance Characteristics

Performance Characteristics
Method Comparison
I The plots and table show the results of a comparison of samples analyzed on the VITROS DT60 II System with those analyzed using the Fiske and Subbarow comparative method2, modified by Dryer, Tamnes, and Routh.11 Testing followed NCCLS Protocol EP9.15

Method Comparison for PHOS DT: Serum Conventional Units


14SI Units

la|

ics'
6-

Q CO

O 2 .

0
0 10 12 14
Comparative Method: Modified Fiske and Subbarow (mmol/L) Comparative Method: Modified Fiske and Subbarow (mg/dL)

Method Comparison for PHOS DT: Serum


Conventional Units (mg/dL) n DT60 II System vs. comparative method 69 Correlation Slope Coefficient 0.96 0.982 Range of Sample Cone. Intercept 0.22 Sy.x 0.63 SI Units (mmol/L) Range of Sample Cone. 0.43-4.08 Intercept 0.07 Sy.x 0.20

1.3-12.6

Precision
Precision was evaluated with quality control materials on VITROS the DT60/DT60 II System following NCCLS Protocol EP5.16 The data presented are a representation of test performance and are provided as a guideline. Variables such as sample handling and storage, reagent handling and storage, laboratory environment, and system maintenance can affect reproducibility of test results.

Precision for PHOS DT: Serum


Conventional Units (mg/dL) Mean Within Within Cone. Day SD* Lab SD** 3.2 0.04 0.08 7.0 0.07 0.14 SI Units (mmol/L) Mean Within Within Cone. Day SD* Lab SD** 1.03 0.01 0.03 2.27 0.02 0.05 Within Lab CV%** 2.6 2.0 No. Observ. 84 84 No. Days 21 21

System VITROS DT60 II

Within Day precision was determined using two runs/day with two replications. Within Lab precision was determined using a single lot of slides and calibrating weekly.

Pub. No. C-350

Version 1.0

INSTRUCTIONS FOR USE


References

PHOSDT
Phosphorus

References
1. 2. 3. 4. 5. 6. 7. 8. 9. 10. Tietz NW (ed). Fundamentals of Clinical Chemistry, ed. 3. Philadelphia: WB Saunders; 706-716; 1987. Fiske CH, Subbarow Y. The Colorimetric Determination of Phosphorus. J. Biol. Chem. 66:375; 1925. Gomori G. A Modification of the Colorimetric Phosphorus Determination for Use with a Photoelectric Colorimeter. J. Lab. Clin. Med.

27:955; 1942.
NCCLS. Protection of Laboratory Workers from Instrument Biohazards and Infectious Diseases Transmitted by Blood, Body Fluids and Tissue; Approved Guideline. NCCLS Document M29 (OSBN 1-56238). NCCLS, Wayne, PA 19087; 1997. Doumas BT, et al. Differences Between Values for Plasma and Serum in Tests Performed in the Ektachem 700 XR Analyzer, and Evaluation of "Plasma Separator Tubes (PST)." Clin. Chem. 35:151-153; 1989. Calam RR. Specimen Processing Separator Gels: An Update. J Clin Immunoassay. 11:86-90; 1988. Tietz NW (ed). Textbook of Clinical Chemistry, ed. 3. Philadelphia: WB Saunders; 1407; 1999. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Venipuncture. NCCLS Document H3. Wayne, PA: NCCLS;

1991.
NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Skin Puncture. NCCLS Document H4. Wayne, PA: NCCLS;

1991.
Clinical Laboratory Handbook for Patient Preparation and Specimen Handling. Fascicle VI: Chemistry/Clinical Microscopy. Northfield, IL: College of American Pathologists; 1992. 11. Dryer RL, Tamnes AR, Routh JL. The Determination of Phophorous and Phosphatase with N-Phenyl-p-phenylenediamine. J. Biol. Chem. 222:177; 1957. AACC Proposed Selected Method. 12. NCCLS. Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition. NCCLS Document C24. Wayne, PA: NCCLS; 1999. 13. Young DS. Effects of Drugs on Clinical Laboratory Tests, ed. 4. Washington D C : AACC Press; 1995. 14. Friedman RB, Young DS. Effects of Disease on Clinical Laboratory Tests. Washington, D C : AACC Press; 1990. 15. NCCLS. Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline. NCCLS Document EP9. Wayne, PA: NCCLS; 1995. 16. NCCLS. User Evaluation of Precision Performance with Clinical Chemistry Devices. NCCLS Document EP5. Wayne, PA: NCCLS; 1992.

Glossary of Symbols
The Glossary of Symbols table defines the symbols used on the VITROS Chemistry Products packaging and on the VITROS Chemistry Products Assay Sheets.

Glossary of Symbols
Do Not Reuse Use by or Expiration Date (YYYY-MM-DD) Lot Number Manufacturer's Serial Number Catalog Number or Product Code Attention: See Instructions for Use. Manufacturer Authorized Representative Contains Sufficient for "n" Tests For In Vitro Diagnostic Use Store At or Below Store At or Above Store Between Consult Instructions for Use Fragile, Handle with Care. Keep Dry This end up

Version 1.0

Pub. No. C-350

PHOSDT
Phosphorus

INSTRUCTIONS FOR USE


Revision History

Revision History
Date of Revision 2003-04-30 Version 1.0 Description of Technical Changes* New format New organization and sections consistent with IVD Directive Specimens Not Recommended - added "Do not use hemolyzed specimens." Specimen Storage and Stability - updated data Materials Required But Not Provided - updated materials > Quality Control Material Selection - added statement regarding ethylene glycol > Method Comparison - updated data and plots > Precision - updated data > References - added all except 2, 3, 8

The change bars indicate the position of a technical amendment to the text with respect to the previous version of the document.

When this Instructions For Use is replaced, sign and date below and retain as specified by local regulations or laboratory policies, as appropriate.

Signature

Obsolete Date

C
EC I REP

Ortho-Clinical Diagnostics Mandeville House 62 The Broadway Amersham Buckinghamshire HP7 OHJ England Ortho-Clinical Diagnostics, Inc. 100 Indigo Creek Drive Rochester, NY 14626-5101

'Ortho-Clinical Diagnostics

VITROS is a trademark of Ortho-Clinical Diagnostics, Inc. Ortho-Clinical Diagnostics, Inc., 2003. All rights reserved. Printed in USA.

Pub. No. C-350

Version 1.0

Products

VITRCD5
C hem i s t r y I

INSTRUCTIONS FOR USE


VITROS Chemistry Products TBIL DT Slides Intended Use

TBIL DT
Total Biiirubin

For in vitro diagnostic use only. VITROS TBIL DT Slides quantitatively measure total biiirubin (TBIL) concentration in serum and plasma.

Summary and Explanation of the Test


Total biiirubin in serum and plasma is the sum of unconjugated biiirubin (Bu), mono- and di-glucuronide conjugated biiirubin (Be), and delta biiirubin (DELB), a biiirubin fraction covalently bound to albumin. With the exception of anicteric jaundice, total serum biiirubin is invariably increased in jaundice. Causes of jaundice are prehepatic, resulting from various hemolytic diseases; hepatic, resulting from hepatocellular injury or obstruction; and posthepatic, resulting from obstruction of the hepatic or common bile ducts.'

Principles of the Procedure


The VITROS TBIL DT Slide method is performed using the VITROS TBIL DT Slide and the VITROS Chemistry Products DT Calibrator Kit on VITROS DT60/DT60 II Chemistry Systems. The VITROS TBIL DT Slide is a multilayered, analytical element coated on a polyester support. The analysis is based on a modification of the classic diazo reaction.2 A drop of patient sample is deposited on the slide and is evenly distributed by the spreading layer to the underlying layers. This layer provides a reflective background for measuring the diazo products of biiirubin and contains all reagents necessary to determine total biiirubin. The method uses dyphylline to dissociate unconjugated biiirubin from albumin. Unconjugated biiirubin, conjugated biiirubin, and albumin-linked biiirubin (delta) subsequently react with the diazonium salt 4-(A/-carboxymethylsulfonyl) benzenediazonium hexafluorophosphate to produce azobilirubin chromophores that have similar molar absorptivities. The change in reflection density is proportional to the total biiirubin concentration present in the sample.

Reaction Sequence
total biiirubin (Bu, Be, and DELB)
dyphylline [4-(A/-carboxymethylsulfonyl)-benzenediazonium hexafluorophosphate]

azobilirubin chromophores

Test Type and Conditions


Test Type and Conditions for TBIL DT
Test Type Colorimetric VITROS DT60/DT60 II Module DT60/DT60 II Approximate Incubation Time 5 minutes Temperature 37C (98.6F) Wavelength 555 nm Sample Drop Volume 10 uL

Warnings and Precautions


For in vitro diagnostic use only. Take care when handling materials and samples of human origin. Since no test method can offer complete assurance that infectious agents are absent, consider all clinical specimens, controls, and calibrators potentially infectious. Handle specimens, solid and liquid waste, and test components in accordance with local regulations and NCCLS Guideline M293 or other published biohazard safety guidelines. For specific warnings and precautions for calibrators, quality control materials, and other components, refer to the instructions for use for the appropriate VITROS product, or to other manufacturer's product literature.

Version 1.0

Pub. No. C-305

TBILDT
Total Bilirubin

INSTRUCTIONS FOR USE


Reagents

Reagents
Slide Diagram

Slide Ingredients Reactive ingredients per cm


2

Dyphylline 0.5 mg and 4-(N-carboxymethylaminosulfonyl) benzene diazonium hexafluorophosphate 57 ^g.

Other ingredients
Pigment, binders, buffer, mordant, surfactants and stabilizer.

1. Upper slide mount 2. Spreading layer (BaSO4) dyphylline diazonium salt 3. Reagent layer buffer, pH 3.0 4. Support layer 6. Lower slide mount

. 5

Slide Handling
Do not use slides with damaged or incompletely sealed packaging.

Slide Preparation
IMPORTANT: The slide must reach room temperature, 18 "-2B C (64 -82 F), before the wrapper is opened. Do not use unopened slides that have been at room temperature, 18 -28 C (64 -82 F) for >48 hours. 1. 2. Remove the unopened slide from the box. Warm the unopened slide for 15 minutes. Unopened slides that remain in the sealed wrapper may be returned to refrigerator or freezer storage. 3. Remove the packaging and place the slide into the loading station within 15 minutes after opening.

Slide Storage and Stability


VITROS TBIL DT Slides are stable until the expiration date on the carton when they are stored and handled as specified.

Slide Storage and Stability for TBIL DT


Slides Unopened Opened Storage Condition Room temperature 18-28C(64 -82 F) Frozen <-18C (<0F Room temperature 18-28C(64 -82F) Stability <48 hours Until expiration date <15 minutes

Specimen Requirements
Handle specimens as biohazardous material. Specimens Recommended
Serum Plasma:4 Heparin Do not use this slide for specimens from neonatal patients less than 14 days old. Refer to "Limitations of the Procedure." Certain collection devices have been reported to affect other analytes and tests.s Confirm that your collection devices are compatible with this test.

NOTE:

IMPORTANT:

Serum and Plasma Specimen Collection and Preparation


Collect specimens using standard laboratory procedures.6 7 Patient Preparation No special patient preparation is necessary.

Pub. No. C-305

Version 1.0

INSTRUCTIONS FOR USE


Testing Procedure

TBIL DT
Total Bilirubin

Special Precautions For the effect of sample hemolysis on test results, refer to "Limitations of the Procedure." Protect specimens from light. Direct exposure to sunlight is reported to cause as much as a 50% loss of serum bilirubin in one hour, especially when the specimen is kept in capillary tubes.2 Exposure to normal laboratory light can result in a significant loss of serum bilirubin after two to three hours. Centrifuge specimens and remove the serum from the cellular material within 4 hours of collection.8 Specimen Handling and Storage WfiMUM(> Handle specimens as biohazardous material.

Handle and store specimens in stoppered containers to avoid contamination and evaporation. Mix samples by gentle inversion and bring to room temperature, 18-28C (64-82F), prior to analysis.

Specimen Storage and Stability for TBIL DT: Serum8


Storage Room temperature Refrigerated Frozen Temperature

18-28C(64-82F) 2-8C (36-46F) <-18C(<0F)

Stability <4 hours <7 days <6 months

Testing Procedure
Materials Provided
VITROS Chemistry Products TBIL DT Slides

Materials Required But Not Provided


| VITROS Chemistry Products DT Calibrator Kit Quality control materials, such as VITROS Chemistry Products DT Control I & II VITROS Chemistry Products 7% BSA or reagent-grade water VITROS DT Pipette

Operating Instructions
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System. Bring all fluids and samples to room temperature, 18-28C (64-82F), prior to analysis. IMPORTANT:

Sample Dilution
| If total bilirubin concentrations exceed the system's reportable (dynamic) range: 1. Dilute the sample with VITROS 7% BSA or reagent-grade water. 2. Reanalyze. 3. Multiply the results by the dilution factor to obtain an estimate of the original sample's total bilirubin concentration.

Calibration
Required Calibrators
VITROS Chemistry Products DT Calibrator Kit, bottles 1, 2, and 3

Calibrator Preparation, Handling, and Storage


Refer to the instructions for use for VITROS DT Calibrator Kit.

Calibration Procedure
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

When to Calibrate
Calibrate: When the slide lot number changes. When critical system parts are replaced due to service or maintenance. When government regulations require. - For example, in the USA, CLIA regulations require calibration or calibration verification at least once every six months. Version 1.0 Pub. No. C-305

TBILDT
Total Bilirubin

INSTRUCTIONS FOR USE


Quality Control

The VITROS TBIL DT test may also need to be calibrated: If quality control results are consistently outside acceptable range. After certain service procedures have been performed. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Calculations
Reflectance from the slide is measured at 555 nm after the fixed incubation time. Once a calibration has been performed for each slide lot, total bilirubin concentration in unknown samples can be determined using the software-resident endpoint colorimetric math model and the response obtained from each unknown test slide.

Validity of a Calibration
Calibration parameters are automatically assessed by the VITROS DT60/DT60II Chemistry System against a set of quality parameters resident within the analyzer software. Failure to meet any of the pre-defined quality parameters results in a failed calibration. The calibration report should be used in conjunction with quality control results to determine the validity of a calibration.

Reportable (Dynamic) Range Reportable (Dynamic) Range for TBIL DT


Conventional (mg/dL) SI Units (umol/L) 2-342

0.1-20.0

For out-of-range samples, refer to "Sample Dilution."

Traceability of the Calibration

I
|

Values assigned to the VITROS Chemistry Products DT Calibrator Kit for total bilirubin are traceable to the Certified NIST (National Institute of Standards and Technology) Reference Material, SRM" (Standard Reference Material) 916a. The Ortho-Clinical Diagnostics (OCD) calibration laboratory uses SRM 916a to calibrate the NCCLS credentialed Jendrassik-Grof method9 to support total bilirubin value assignment for the VITROS DT Calibrator Kit.

Quality Control
Procedure Recommendations
WARNING: Handle quality control materials as biohazardous material. Choose control levels that check the clinically relevant range. Analyze quality control materials in the same manner as patient samples, before or during patient sample processing. To verify system performance, analyze control materials: - After calibration. - According to local regulations or at least once each day that the test is being performed. - After specified service procedures are performed. Refer to the operator's manual for your VITROS DT60/DT60 II System. If control results fall outside your acceptable range, investigate the cause before deciding whether to report patient results. For general quality control recommendations, refer to Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition'10 or other published guidelines. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Quality Control Material Selection


I | IMPORTANT; VITROS DT Control I & II are recommended for use with the VITROS DT60/DT60 II Chemistry System. Evaluate the performance of other commercial control fluids for compatibility with this test before using for quality control.

Control materials other than VITROS DT Controls I & II may show a difference when compared with other total bilirubin methods if they: - Depart from a true human matrix. - Contain high concentrations of preservatives, stabilizers, or other nonphysiological additives. Do not use control materials stabilized with ethylene glycol.

Pub. No. C-305

Version 1.0

la

V|T

.S)

INSTRUCTIONS FOR USE


Expected Values and Reporting Units

TBILDT
Total Bilirubin

Quality Control Material Preparation and Storage


Refer to the instructions for use for VITROS Chemistry Products DT Control I & II or to other manufacturer's product literature.

Expected Values and Reporting Units


Reference Interval
This reference interval is the central 95% of results from a study of 110 apparently healthy adults with normal liver enzymes from a working population (85 females and 25 males).

Reference Interval for TBIL DT Conventional Units (mg/dL) 0.2-1.3

SI Units (umol/L) 3-22

Each laboratory should confirm the validity of these intervals for the population it serves.

Reporting Units and Unit Conversion


The VITROS DT60/DT60 II Chemistry System may be programmed to report total bilirubin results in conventional and SI units.

Reporting Units and Unit Conversion for TBIL DT


Conventional Units mg/dL SI Units umol/L (mg/dL x 17.1)

Limitations of the Procedure


Known Interferences
The VITROS TBIL DT Slide method was screened for interfering substances following NCCLS Protocol EP7.11 The following substances, when tested at the concentrations indicated, caused the bias shown.

Known Interfering Substances for TBIL DT


Interferent* Levodopa Interferent Concentration Bilirubin Cone. Conv. (mg/dL) SI (Mmol/L) 0.4 7 (1.52mmol/L)** 17.8 304 (1.52mmol/L)** 0.4 (0.52 mmol/L) 7 (0.52 mmol/L) 17.6 301 (0.3 mmol/L) 0.4 7 (0.3 mmol/L) 17.4 298 0.3 5 (69 umol/L) 16.4 (69 umol/L) 280 0.5 9 (1 g/L) 0.5 9 (2 g/L) 9 0.5 (4 g/L) 10.5 180 (1 g/L) 10.5 180 (2 g/L) 10.5 180 (4 g/L) 15.0 257 (1 g/L) 15.0 257 (2 g/L) 15.0 257 (4 g/L) Bias Conv. (mg/dL) SI +1.4 -9.4 +0.4 +2.5 +3.8 +3.5 +0.5 +0.7 +0.1 +0.4 +1.3 -1.0 -1.2 -1.0 -1.2 -1.7 -1.9 (Mmol/L) +24 -161 +7 +43 +65 +60 +9 +12 +2 +7 +22 -17 -21 -17 -21 -29 -32

300 ng/mL** 300 ug/mL** 8 mg/dL 4-Aminosalicylic acid 8 mg/dL Phenazopyridine 8 mg/dL 8 mg/dL Biliverdin 4 mg/dL 4 mg/dL 100 mg/dL Hemoglobin 200 mg/dL 400 mg/dL 100 mg/dL 200 mg/dL 400 mg/dL 100 mg/dL 200 mg/dL . 400 mg/dL
* **

It is possible that other interfering substances may be encountered. These results are representative; however, your results may differ somewhat due to test-to-test variation. The degree of interference at concentrations other than those listed might not be predictable. This concentration may be present in the treatment of Parkinson's disease.

Other Limitations
Accuracy of VITROS TBIL DT Slides results on specimens from neonates less than 14 days old has not been demonstrated. Therefore, these specimens should be analyzed using VITROS NBIL DT Slides. It is important to maintain consistency in bilirubin methodology. If VITROS NBIL DT Slides are initially used to monitor a patient, continue to monitor that patient with VITROS NBIL DT Slides. Do not switch to VITROS TBIL DT Slides regardless of the patient's age. Results from the VITROS TBIL DT Slide may not be accurate at elevation greater than 6000 feet (approx. 1800 meters) above sea level. Pub. No. C-305

Version 1.0

VITROS 0

TBIIDT
Total Bilirubin

INSTRUCTIONS FOR USE


Performance Characteristics
Cefotiam (Pansporin) has been reported to show very large positive biases on VITROS TBIL DT Slide results.12 This drug is normally cleared through the kidney. Biases will be largest in specimens from patients with renal insufficiency and may be as large as 5 mg/dL (86 (.imol/L). Drugs and other compounds that are diazo-reactive or that absorb light in the vicinity of 555 nm may interfere. Certain drugs and clinical conditions are known to alter total bilirubin concentration in vivo. For additional information, refer to one of the published summaries.1314

Performance Characteristics
Method Comparison

The plots and table show the results of a comparison of samples analyzed on the VITROS DT60 II System with those analyzed using the Jendrassik-Grof comparative method, Doumas modification.9

Method Comparison for TBIL DT: Serum


Conventional Units

400 y =x

10

15

20

200

300

400

Comparative Method: Jendrassik-Grof Doumas Modification (mg/dL)

Comparative Method: Jendrassik-Grof Doumas Modification (Mmol/L)

Method Comparison for TBIL DT: Serum


Conventional Units (mg/dL) n DT60 II System vs. comparative method 67 Correlation Slope Coefficient 0.99 0.998 Range of Sample Cone. 0.1-17.8 Intercept Sy.x 0.02 0.31 SI Units (Mmol/L) Range of Sample Cone. 2-304 Intercept 0.38 Sy.x 5.30

Pub. No. C-305

Version 1.0

El VtTRtCpS

INSTRUCTIONS FOR USE


References Precision

TBILDT
Total Bilirubin

Precision was evaluated with quality control materials on the VITROS DT60 II System following NCCLS Protocol EP5.15 The data presented are a representation of test performance and are provided as a guideline. Variables such as sample handling and storage, reagent handling and storage, laboratory environment, and system maintenance can affect reproducibility of test results. Precision for TBIL DT: Serum Conventional Units (mg/dL) System VITROS DT60 II Mean Cone. 1.3 13.3 Within Day SD* 0.06 0.18 Within Lab SD** 0.07 0.23 Mean Cone. 23 227 SI Units (umol/L) Within Day SD* 1.0 3.1 Within Lab SD** 1.2 3.9 Within Lab CV%** 5.1 1.7 No. Observ. 88 88 No. Days 22 22

Within Day precision was determined using two runs/day with two replications. Within Lab precision was determined using a single lot of slides and calibrating weekly.

References
Tietz NW (ed). Fundamentals of Clinical Chemistry, ed. 3. Philadelphia: WB Saunders; 730-736; 1987. Routh Jl: Liver Function. In Tietz NW(ed). Fundamentals of Clinical Chemistry, ed 2. Philadelphia; WB Saunders; 1037; 1976. NCCLS. Protection of Laboratory Workers from Instrument Biohazards and Infectious Diseases Transmitted by Blood, Body Fluids and Tissue; Approved Guideline. NCCLS Document M29 (OSBN 1-56238). NCCLS, Wayne, PA 19087; 1997. 4. Doumas BT, et al. Differences Between Values for Plasma and Serum in Tests Performed in the Ektachem 700 XR Analyzer, and Evaluation of "Plasma Separator Tubes (PST)." Clin. Chem. 35:151-153; 1989. 5. Calam RR. Specimen Processing Separator Gels: An Update. J Clin Immunoassay. 11:86-90; 1988. 6. NCCLS. Procedures forthe Collection of Diagnostic Blood Specimens by Venipuncture. NCCLS Document H3. Wayne, PA: NCCLS; 1991. 7. NCCLS. Procedures forthe Collection of Diagnostic Blood Specimens by Skin Puncture. NCCLS Document H4. Wayne, PA: NCCLS; 1991. 8. Clinical Laboratory Handbook for Patient Preparation and Specimen Handling. Fascicle VI: Chemistry/Clinical Microscopy. Northfield, IL: College of American Pathologists; 1992. 9. Doumas B.T., Bayse D.D., et al. A candidate reference method for determination of bilirubin in serum. Test for transferability. Clin. Chem. 29; 297-301; 1983. 10. NCCLS. Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition. NCCLS Document C24. Wayne, PA: NCCLS; 1999. 11. NCCLS. Interference Testing in Clinical Chemistry. NCCLS Document EP7. Wayne, PA: NCCLS; 1986. 12. Pickert A, Riedlinger I, Stumvill M. Interference of Cefotiam with Total Bilirubin Measured with the Ektachem Analyzer. Clin. Chem. 38:599-600; 1992. 13. Young DS. Effects of Drugs on Clinical Laboratory Tests, ed. 4. Washington D.C.: AACC Press; 1995. 14. Friedman RB, Young DS. Effects of Disease on Clinical Laboratory Tests. Washington, D C : AACC Press; 1990. 15. NCCLS. User Evaluation of Precision Performance with Clinical Chemistry Devices. NCCLS Document EP5. Wayne, PA: NCCLS; 1992. 1. 2. 3.

Glossary of Symbols
The Glossary of Symbols table defines the symbols used on the VITROS Chemistry Products packaging and on the VITROS Chemistry Products Assay Sheets.

Glossary of Symbols
Do Not Reuse Use by or Expiration Date (YYYY-MM-DD) Lot Number Manufacturer's Serial Number Catalog Number or Product Code Attention: See Instructions for Use. Manufacturer Authorized Representative Contains Sufficient for "n" Tests For In Vitro Diagnostic Use Store Between

m
i

Consult Instructions for Use Fragile, Handle with Care. Keep Dry This end up

Store At or Below Store At or Above

if

Version 1.0

Pub. No. C-305

TBILDT
Total Bilirubin

INSTRUCTIONS FOR USE


Revision History

Revision History
Date of Revision 2003-10-01 Version 1.0 Description of Technical Changes* New format New organization and sections consistent with IVD Directive Materials Required But Not Provided and Sample Dilution - added VITROS 7% BSA Reference Interval - updated all data Known interfering Substances - updated values; added hemoglobin; changed 5-aminosalicylic acid to 4-aminosalicylic acid Method Comparison - updated all comparisons and the plots Precision - updated all values References - added all except 6,13,14

The change bars indicate the position of a technical amendment to the text with respect to the previous version of the document.

When this Instructions For Use is replaced, sign and date below and retain as specified by local regulations or laboratory policies, as appropriate.

Signature

Obsolete Date

C
REP

Ortho-Clinical Diagnostics Mandeville House 62 The Broadway Amersham Buckinghamshire HP7 OHJ England Ortho-Clinical Diagnostics, Inc. 100 Indigo Creek Drive Rochester, NY 14626-5101

QihoClinical Diagnostics
company

VITROS is a trademark of Ortho-Clinical Diagnostics, Inc. Ortho-Ciinical Diagnostics, Inc., 2003. All rights reserved. Printed in USA.

Pub. No. C-305

Version 1.0

[ Prodwcts

VITRII15
Chemistry!

INSTRUCTIONS FOR USE


VITROS Chemistry Products THEO DT Slides Intended Use

THEO OT
Theophylline

For in vitro diagnostic use only. VITROS THEO DT Slides quantitatively measure theophylline (THEO) concentration in serum and plasma.

Summary and Explanation of the Test


Theophylline is a drug used in the treatment of asthma. Theophylline is an effective bronchodilator. The therapeutic and toxic effects of theophylline are related to the plasma concentrations of the drug. Theophylline measurements are used to monitor patient compliance and therapy and to diagnose potential overdose. Toxic effects include nausea, vomiting, diarrhea, headache, tachycardia, arrhythmias, and convulsions.'

Principles of the Procedure


The VITROS THEO DT Slide method is performed using the VITROS THEO DT Slide and the VITROS Chemistry Products DT Specialty Calibrator Kit on VITROS DT60/DT60 II Chemistry Systems. The VITROS THEO DT Slide is a multilayered, analytical element coated on a polyester support. The assay is based on the inhibition of beef liver alkaline phosphatase (ALKP) activity by theophylline, which acts as a potent uncompetitive inhibitor of this isoenzyme. A drop of patient sample is deposited on the slide and is evenly distributed by the spreading layer to the underlying layers. p-Nitrophenyl phosphate (PNPP) migrates from the spreading layer to the reagent layer. If no theophylline is present, the alkaline phosphatase reaction proceeds at its maximum rate. If theophylline is present, it uncompetitively inhibits the beef liver isoenzyme, and the reaction rate is reduced. The assay is performed at pH 8.5 so that samples containing endogenous alkaline phosphatase (maximum activity at pH 10.5) will not interfere. The product, p-nitrophenol, remains in the transparent reagent layer. The rate of change in reflection density is proportional to the theophylline concentration in the sample.

Reaction Sequence
p-nitrophenyl phosphate
beef liver ALKP MgCI2

- > p-nitrophenol + H3PO4

Test Type and Conditions


Test Type and Conditions for THEO DT
Test Type Rate VITROS DT60/DT60 II Module DTSC/DTSC II Approximate Incubation Time 2 minutes Temperature 37C (98.6F) Wavelength 400 nm Sample Drop Volume 10 ML

Warnings and Precautions


For in vitro diagnostic use only. Take care when handling materials and samples of human origin. Since no test method can offer complete assurance that infectious agents are absent, consider all clinical specimens, controls, and calibrators potentially infectious. Handle specimens, solid and liquid waste, and test components in accordance with local regulations and NCCLS Guideline M292 or other published biohazard safety guidelines. For specific warnings and precautions for calibrators, quality control materials, and other components, refer to the instructions for use for the appropriate VITROS product, or to other manufacturer's product literature.

Version 1.0

Pub.

No. C-347

THEODT
Theophylline

INSTRUCTIONS FOR USE


Reagents

Reagents
Slide Diagram

Slide Ingredients Reactive ingredients per cm


2

1 , 2 . 3

Alkaline phosphatase (beef liver, E.C.3.1.3.1) 0.02 U; magnesium chloride 3 ug; and p-nitrophenyl phosphate 0.3 mg.

Other ingredients
Pigment, binders, surfactant, buffer, enzyme cofactor, protein, stabilizer and cross-linking agent.
- $

1. Upper slide mount 2. Spreading layer (BaSO4) p-nitrophenyl phosphate 3. Reagent layer ALKP buffer, pM 8.5 4. Support layer 5. Lower slide mount

Slide Handling
Do not use slides with damaged or incompletely sealed packaging.

Slide Preparation
IMPORTANT: The slide must reach room temperature, 18-28 C (64 -82 F), before the wrapper is opened. Do not use unopened slides that have been at room temperature, 18 - 2 8 'C (64 -82 F) for >48 hours. 1. Remove the unopened slide from the box. 2. Warm the unopened slide for 15 minutes. Unopened slides that remain in the sealed wrapper may be returned to refrigerator or freezer storage. 3. Remove the packaging and place the slide into the loading station within 15 minutes after opening.

Slide Storage and Stability


VITROS THEO DT Slides are stable until the expiration date on the carton when they are stored and handled as specified.

Slide Storage and Stability for THEO DT Slides Storage Condition


Unopened Opened Room temperature Frozen Room temperature 18-28C (64-82F) <-18C (<0F) 18-28C (64-82F)

Stability <48 hours Until expiration date <15 minutes

Specimen Requirements
Handle specimens as biohazardous material.

Specimens Recommended
Serum Plasma: Heparin Certain collection devices have been reported to affect other analytes and tests.3 Confirm that your collection devices are compatible with this test.

IMPORTANT:

Specimens Not Recommended


Plasma 4 Substances that chelate magnesium such as: EDTA Citrate Fluoride oxalate

Serum and Plasma


Specimen Collection and Preparation
Collect specimens using standard laboratory procedures.56 Patient Preparation No special patient preparation is necessary.

Pub. No. C-347

Version 1.0

INSTRUCTIONS FOR USE


Testing Procedure Special Precautions Centrifuge specimens and remove the serum from the cellular material within 4 hours of collection.7 Storage

THEO DT
Theophylline

Specimen Handling and

Handle specimens as biohazardous material. Handle and store specimens in stoppered containers to avoid contamination and evaporation. Mix samples by gentle inversion and bring to room temperature, 18-28C (64-82F), prior to analysis. Specimen Storage and Stability for THEO DT: Serum and Plasma 7 Storage Room temperature Refrigerated Frozen Temperature 18-28C(64-82F) 2-8C (36-46F) <-18C(<0F) Stability <7 days <7 days <60 days

Testing Procedure
Materials Provided
VITROS Chemistry Products THEO DT Slides

Materials Required But Not Provided


| VITROS Chemistry Products DT Specialty Calibrator Kit Quality control materials, such as VITROS Chemistry Products DT Control I & II VITROS Chemistry Products 7% BSA or reagent-grade water VITROS DT Pipette

Operating Instructions
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System. Bring all fluids and samples to room temperature, 18-28C (64-82F), prior to analysis. IMPORTANT:

Sample Dilution
I If theophylline concentrations exceed the system's reportable (dynamic) range: 1. Dilute the sample with VITROS 7% BSA or reagent-grade water. 2. Reanalyze. 3. Multiply the results by the dilution factor to obtain an estimate of the original sample's theophylline concentration.

Calibration
Required Calibrators
VITROS Chemistry Products DT Specialty Calibrator Kit, bottles 1, 2, and 4

Calibrator Preparation, Handling, and Storage


Refer to the instructions for use for VITROS DT Specialty Calibrator Kit.

Calibration Procedure
Refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Version 1.0

Pub. No. C-347

THEODT
Theophylline

INSTRUCTIONS FOR USE


Quality Control

When to Calibrate
Calibrate: When the slide lot number changes. ' When critical system parts are replaced due to service or maintenance. When government regulations require. - For example, in the USA, CLIA regulations require calibration or calibration verification at least once every six months. The VITROS THEO DT test may also need to be calibrated: If quality control results are consistently outside acceptable range. After certain service procedures have been performed. For additional information, refer to the operator's manual for your VITROS DT60/DT60II Chemistry System.

Calculations
Reflectance from the slide is read at 400 nm during the incubation period, and the rate of change in reflectance is calculated. Once a calibration has been performed for each slide lot, theophylline concentration in unknown samples can be determined using the software-resident rate math model and the change in reflectance calculated for each unknown test slide.

Validity of a Calibration
Calibration parameters are automatically assessed by the VITROS DT60/DT60II Chemistry System against a set of quality parameters resident within the analyzer software. Failure to meet any of the pre-defined quality parameters results in a failed calibration. The calibration report should be used in conjunction with quality control results to determine the validity of a calibration.

Re portable (Dynamic) Range Reportable (Dynamic) Range for THEO DT


Conventional Units (ug/mL) SI Units (umol/L) 5.6-222.0

1.0-40.0

For out-of-range samples, refer to "Sample Dilution."

Traceability of the Calibration


Values assigned to the VITROS Chemistry Products DT Specialty Calibrator Kit for theophylline are traceable to gravimetrically prepared standards made from reagent grade theophylline. The Ortho-Clinical Diagnostics calibration laboratory uses the gravimetrically prepared standard to calibrate an HPLC (High Performance Liquid Chromatography) method for theophylline8 to support value assignment for the VITROS DT Specialty Calibrator Kit.

Quality Control
Procedure Recommendations
| , <. Handle quality control materials as biohazardous material. Choose control levels that check the clinically relevant range. Analyze quality control materials in the same manner as patient samples, before or during patient sample processing. . To verify system performance, analyze control materials: - After calibration. - According to local regulations or at least once each day that the test is being performed. - After specified service procedures are performed. Refer to the operator's manual for your VITROS DT60/DT60 II System. If control results fall outside your acceptable range, investigate the cause before deciding whether to report patient results. For general quality control recommendations, refer to Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition9 or other published guidelines. For additional information, refer to the operator's manual for your VITROS DT60/DT60 II Chemistry System.

Pub.

No. C-347

Version 1.0

INSTRUCTIONS FOR USE


Expected Values and Reporting Units

THEO DT
Theophylline

Quality Control Material Selection


VITROS DT Control I & II are recommended for use with the VITROS DT60/DT60 II Chemistry System. Evaluate the performance of other commercial control fluids for compatibility with this test before using for quality control. Control materials other than VITROS DT Controls I & II may show a difference when compared with other theophylline methods if they: - Depart from a true human matrix. - Contain high concentrations of preservatives, stabilizers, or other nonphysiological additives. Quality-control and proficiency survey specimens that contain salicylate may show a positive bias compared with their target theophylline concentrations. Refer to "Limitations of the Procedure." Do not use control materials stabilized with ethylene glycol.

Quality Control Material Preparation and Storage


Refer to the instructions for use for VITROS Chemistry Products DT Control I & I or to other manufacturer's product literature.

Expected Values and Reporting Units


Reference Interval
This reference interval is based on an external study.10

Reference Interval for THEO DT


Conventional Units (ug/mL) Therapeutic range

10.0-20.0

SI Units (umol/L) 55.5-111.0

Each laboratory should confirm the validity of these intervals for the population it serves. In some cases the most effective therapeutic concentration may be outside this range. Monitor patients for efficacy of treatment and for adverse symptoms.

Reporting Units and Unit Conversion


The VITROS DT60/DT60 II Chemistry System may be programmed to report theophylline results in conventional and SI units.

Reporting Units and Unit Conversion for THEO DT


Conventional Units ug/mL SI Units umol/L (ug/mL x 5.55)

Limitations of the Procedure


Known Interferences
Uremic specimens have shown positive biases of 3-5 ug/mL (17-28 umol/L) in the therapeutic range for theophylline. The VITROS THEO DT Slide method was screened for interfering substances following NCCLS Protocol EP7.1' The substances listed in the table, when tested at the concentrations indicated, caused the bias shown.

Known Interfering Substances for THEO DT


Interferent* Salicylate Alkaline Phosphatase
*

Interferent Concentration 30mg/dL (2.17 mmol/L) 700 U/L

Conv. (ug/mL) 20 20

Theophylline Concentration SI (umol/L) 111 111

Bias Conv. (ug/mL) SI (pmol/L) 3.0 16.7 2.0 11.1

It is possible that other interfering substances may be encountered. These results are representative; however, your results may differ somewhat due to test-to-test variation. The degree of interference at concentrations other than those listed might not be predictable.

Other Limitations
Certain drugs and clinical conditions are known to alter theophylline concentration in vivo. For additional information, refer to one of the published summaries.1213

Version 1.0

Pub. No. C-347

THEO DT
Theophylline

INSTRUCTIONS FOR USE


Performance Characteristics

Performance Characteristics
Method Comparison

The plots and table show the results of a comparison of samples analyzed on the VITROS DT60 II System with those analyzed using the VITROS 950 System. Testing followed NCCLS Protocol EP9.14 Method Comparison for THEO DT: Serum Conventional Units
.?u

SI Units
y=X

y =x

250
200

40 ?0 20 100 (0 20 30 40 50
Comparative Method: VITROS 950 System (Mg/mL)

J*
Q

150 100

so
0 50
100

150

200

250

Comparative Method: VITROS 950 System ((Jmol/L)

Method Comparison for THEO DT: Serum


Conventional Units (ug'mL) n DT60 II System vs. 950 System 60 Correlation Slope Coefficient 0.97 0.991 Range of Sample Cone. Intercept 1.31 Sy.x 1.37 SI Units (umol/L) Range of Sample Cone. Intercept 7.27 Sy.x 7.62

0.6-37.9

3.5-210.5

Precision
Precision was evaluated with quality control materials on the VITROS DT60 II System following NCCLS Protocol EP5.15 The data presented are a representation of test performance and are provided as a guideline. Variables such as sample handling and storage, reagent handling and storage, laboratory environment, and system maintenance can affect reproducibility of test results.

Precision for THEO DT: Serum


Conventional Units (ug/mL) System VITROS DT60 II Mean Cone. 12.9 21.7 Within Day SD* 0.49 0.67 Within Lab SD** 0.65 0.93 SI Units (umol/L) Mean Cone. 71.3 120.2 Within Day SD* 2.74 3.73 Within Lab SD** 3.63 5.16 Within Lab CV%** 5.1 4.3 No. Observ. 88 88 No. Days 22 22

Within Day precision was determined using two runs/day with two replications. Within Lab precision was determined using a single lot of slides and calibrating weekly.

Pub. No. C-347

Version 1.0

INSTRUCTIONS FOR USE


References

THEO DT
Theophylline

References
1. 2. 3. 4. 5. 6. Tietz NW (ed). Fundamentals of Clinical Chemistry, ed. 3. Philadelphia: WB Saunders; 859; 1987. NCCLS. Protection of Laboratory Workers from Instrument Biohazards and Infectious Diseases Transmitted by Blood, Body Fluids and Tissue; Approved Guideline. NCCLS Document M29 (OSBN 1-56238). NCCLS, Wayne, PA 19087; 1997. Calam RR. Specimen Processing Separator Gels: An Update. J Clin Immunoassay. 11:86-90; 1988. Kaplan L, PesceA. Clinical Chemistry: Theory, Analysis, and Correlation, ed. 2. CVMosby; 900; 1989. NCCLS. Procedures for the Collection of Diagnostic Blood Specimens by Venipuncture. NCCLS Document H3. Wayne, PA: NCCLS;

1991.
NCCLS. Procedures for the. Collection of Diagnostic Blood Specimens by Skin Puncture. NCCLS Document H4. Wayne, PA: NCCLS; 1991. 7. Clinical Laboratory Handbook for Patient Preparation and Specimen Handling. Fascicle IV: Therapeutic Drug Monitoring/Toxicology. Skokie, IL: College of American Pathologists; 1985. 8. Lauff J. A Reference Procedure for the Determination of Theophylline and Related Xanthines in Serum by Dynamic Ion-Exchange HPLC. J. Chrom. 417:99-109; 1987. 9. NCCLS. Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline-Second Edition. NCCLS Document C24. Wayne, PA: NCCLS; 1999. 10. Hendele L, Weinberger MM. Theophylline Therapeutic Use and Serum Concentration Monitoring, in Taylor WJ and Finn AL (eds). Individualizing Drug TherapyPractical Applications of Drug Monitoring. New York: Gross, Townsend, Frank; 1:31-65; 1981. 11. NCCLS. Interference Testing in Clinical Chemistry. NCCLS Document EP7. Wayne, PA: NCCLS; 1986. 12. Young DS. Effects of Drugs on Clinical Laboratory Tests, ed. 4. Washington D.C.: AACC Press; 1995. 13. Friedman RB, Young DS. Effects of Disease on Clinical Laboratory Tests. Washington, D.C.: AACC Press; 1990. 14. NCCLS. Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline. NCCLS Document EP9. Wayne, PA: NCCLS; 1995. 15. NCCLS. User Evaluation of Precision Performance with Clinical Chemistry Devices. NCCLS Document EP5. Wayne, PA: NCCLS; 1992.

Glossary of Symbols
The Glossary of Symbols table defines the symbols used on the VITROS Chemistry Products packaging and on the VITROS Chemistry Products Assay Sheets.

Glossary of Symbols
Do Not Reuse Use by or Expiration Date (YYYY-MM-DD) Lot Number Manufacturer's Serial Number Catalog Number or Product Code Attention: See Instructions for Use.

ml
| t t [ m$t> I Vr7 V

Manufacturer Authorized Representative Contains Sufficient for "n" Tests For In Vitro Diagnostic Use Store At or Below Store At or Above

Store Between Consult Instructions for Use Fragile, Handle with Care. Keep Dry This end up

Version 1.0

Pub. No. C-347

VITFJCpS Q

THEODT
Theophylline

INSTRUCTIONS FOR USE


Revision History

Revision History
Date of Revision 2003-10-01 Version 1.0 Description of Technical Changes* New format > New organization and sections consistent with IVD Directive > Materials Required But Not Provided and Sample Dilution - added VITROS 7% BSA; removed isotonic saline > Limitations of the Procedure - updated values > Method Comparison - updated all data and plots Precision - updated all values References - added all

The change bars indicate the position of a technical amendment to the text with respect to the previous version of the document.

When this Instructions For Use is replaced, sign and date below and retain as specified by local regulations or laboratory policies, as appropriate.

Signature

Obsolete Date

C
FU5P

Ortho-Clinical Diagnostics Mandeville House 62 The Broadway Amersham Buckinghamshire HP7 OHJ England Ortho-Clinical Diagnostics, Inc. 100 Indigo Creek Drive Rochester, NY 14626-5101

Qtho-Clinical Diagnostics
company

VITROS is a trademark of Ortho-Clinical Diagnostics, Inc. Ortho-Clinical Diagnostics, Inc., 2003. All rights reserved. Printed in USA.

Pub. No. C-347

Version 1.0

INSTRUCTIONS FOR USE


VITROS Chemistry Products TP DT Slides Intended Use
For in vitro diagnostic use only. VITROS TP DT Slides quantitatively measure total protein (TP) concentration in serum and plasma.

TPDT
Total Protein

Summary and Explanation of the Test


Serum proteins transport drugs and metabolites and maintain plasma osmotic pressure. Most serum proteins are synthesized in the liver, with the exception of gamma globulins. One of the most important serum proteins produced in the liver is albumin. Total serum protein concentration can be used for evaluation of nutritional status. Causes of high total serum protein concentration include dehydration, Waldenstrom's macroglobulinemia, multiple myeloma, hyperglobulinemia, granulomatous diseases, and some tropical diseases. Total protein concentration is occasionally increased in collagen diseases, lupus erythematosus, and other instances of chronic infection or inflammation. Causes of low total serum protein concentration include pregnancy, excessive intravenous fluid administration, cirrhosis or other liver diseases, chronic alcoholism, heart failure, nephrotic syndrome, glomerulonephritis, neoplasia, protein-losing enteropathies, malabsorption, and severe malnutrition.1

Principles of the Procedure


The VITROS TP DT Slide method is performed using the VITROS TP DT Slide and the VITROS Chemistry Products DT Calibrator Kit on VITROS DT60/DT60 II Chemistry Systems. The VITROS TP DT Slide is a muitilayered, analytical element coated on a polyester support. The method of analysis is based on the biuret reaction,2 which produces a violet complex when protein reacts with cupric ion (Cu*2) in an alkaline medium. A drop of patient sample is deposited on the slide and is evenly distributed by the