E

HBsAg (V2)
7A40-22

B7P400
33-0734/R8

HBsAg (V2)
Antibody to Hepatitis B Surface Antigen (Goat):
Conjugate [Mouse Monoclonal Coated Microparticles]

Note Changes Highlighted

1998, 2002 Abbott / AxSYM HBsAg (V2)

November 2002

Key to symbols used

List Number Lot Number

For In Vitro Diagnostic

Use Expiration Date

Store at 2-8C Reagent Pack

Reaction Vessels
Store at 15-30C

Matrix Cells
CAUTION: Handle human
sourced materials as
potentially infectious.
Consult instructions for use. Sample Cups
(Infection Risk)

Consult instructions
Legal for use.
Manufacturer

See REAGENTS section for a full explanation of symbols used in reagent component naming.

ABBOTT
Diagnostics Division
1
NAME 1 Bottle (17.6 mL) Biotinylated Anti-HBs (Goat, IgG) in TRIS buffer
HBsAg - Hepatitis B Surface Antigen containing animal sera (Goat, Calf, Rabbit, Mouse). Minimum
concentration: 0.25 g/mL. Preservatives: Sodium Azide, Nipasept,
INTENDED USE Quinolone. (Reagent Bottle 3)
AxSYM HBsAg (V2) is a third generation microparticle enzyme immuno-
1 Bottle (6 mL) AxSYM HBsAg (V2) Index Calibrator. Recalcified
assay for the qualitative detection of Hepatitis B Surface Antigen (HBsAg) in
human plasma nonreactive for HBsAg, HIV-1 Ag, anti-HBs, anti-HCV and
human serum or plasma.
anti-HIV-1/HIV-2. Preservative: Sodium Azide. Dye: Green (Acid Yellow
SUMMARY AND EXPLANATION OF THE TEST No. 23 and Acid Blue No. 9).
Enzyme immunoassays for the detection of HBsAg were first described by Nipasept is a registered trademark of Nipa Laboratories, Wilmington, Delaware.
Engvall and Perlmann1-3 and VanWeemen and Schuurs4 in 1971. In 1976 CONTROLS
and 1977, solid phase "sandwich" enzyme immunoassays were developed
AxSYM HBsAg Controls (7A40-10)
in which HBsAg was captured on a solid phase coated with polyclonal
antibodies against HBsAg (anti-HBs) and then detected with anti-HBs 2 Bottles (8 mL each) of AxSYM HBsAg Controls are prepared with recalcified
conjugated to an enzyme.5-7 In the early 1980's, monoclonal anti-HBs based human plasma.
assays were developed for the detection of HBsAg.8-13 AxSYM HBsAg (V2) The Negative Control is nonreactive for HBsAg, HIV-1 Ag,
is an enzyme immunoassay which uses microparticles coated with anti-HBs, anti-HCV and anti-HIV-1/HIV-2.
monoclonal anti-HBs for the detection of HBsAg. The Positive Control is reactive for HBsAg and nonreactive for
Assays for HBsAg are used to screen blood and blood products for the HIV-1 Ag, anti-HCV, anti-HBs and anti-HIV-1/HIV-2.
presence of HBsAg to prevent transmission of hepatitis B virus (HBV) to Preservative: Sodium Azide.
recipients of these products. HBsAg assays are also routinely used to
The AxSYM HBsAg Controls have the following ranges:
diagnose suspected HBV infection and to monitor the status of infected
individuals; i.e., whether the patient's infection has resolved or the patient HBsAg Control Control
has become a chronic carrier of the virus.14 In addition, these assays are Concen- Range Range
used to evaluate the efficacy of anti-viral drugs by monitoring the levels of Control Color tration S/N S/CO
HBsAg in patient sera or plasma.15 The prenatal screening of all pregnant Natural 0 ng/mL 0.70 - 1.30 0.35 - 0.65
women has been recommended by the U.S. Centers for Disease Control so Blue* 4 - 15 ng/mL 7.00 - 63.00 3.50 - 31.50
that newborns from HBV carrier mothers may obtain prophylactic
treatment.16,17 * Dye: Bromophenol Blue.
Samples nonreactive by AxSYM HBsAg (V2) are considered negative for The AxSYM HBsAg (V2) reporting unit is factory set to S/N. An alternate
HBsAg and need not be tested further. A reactive sample should be retested unit, S/CO may be selected for reporting results (Assay Parameter 45).
by AxSYM HBsAg (V2) to determine whether it is repeatedly reactive. A
OTHER REAGENTS
sample which is found to be repeatedly reactive should be confirmed by
neutralization procedures utilizing human anti-HBs, such as the AxSYM AxSYM Probe Cleaning Solution (9A35-04/9A35-05)
HBsAg Confirmatory assay. If the sample is neutralized in a confirmatory 4 Bottles (110 mL each)/2 Bottles (220 mL
assay, the sample is considered positive for HBsAg and does not need to be each) AxSYM Probe Cleaning Solution containing 2%
tested further. Tetraethylammoniumhydroxide (TEAH).
BIOLOGICAL PRINCIPLES OF THE PROCEDURE Solution 1 (MUP) (8A47-04)
AxSYM HBsAg (V2) is based on the Microparticle Enzyme Immunoassay
(MEIA) technology. 4 Bottles (230 mL each) Solution 1 (MUP) containing
4-Methylumbelliferyl Phosphate, 1.2 mM, in buffer. Preservative: Sodium
Sample and all AxSYM HBsAg (V2) reagents required for one test are pipetted
Azide.
by the Sampling Probe into various wells of a reaction vessel (RV) in the
Sampling Center. The RV is immediately transferred into the Processing Solution 3 (Matrix Cell Wash) (8A81-04)
Center. Further pipetting is done in the Processing Center by the Processing
4 Bottles (1000 mL each) Solution 3
Probe.
(Matrix Cell Wash) containing 0.3 M Sodium Chloride in TRIS buffer.
The reactions occur in the following sequence: Preservatives: Sodium Azide and Antimicrobial Agents.
Sample, Anti-HBs Coated Microparticles and Biotinylated Anti-HBs are
combined in one RV well. Solution 4 (Line Diluent) (8A46-01)
When HBsAg is present in the sample, it binds to the Anti-HBs Coated 1 Bottle (10 L) Solution 4 (Line Diluent)
Microparticles and Biotinylated Anti-HBs, forming an antibody-antigen- containing 0.1 M Phosphate buffer. Preservatives: Sodium Azide and
antibody complex in the reaction mixture. Antimicrobial Agent.
A portion of the reaction mixture is transferred to the matrix cell. The
microparticles bind irreversibly to the glass fiber matrix. WARNINGS AND PRECAUTIONS
The Anti-Biotin:Alkaline Phosphatase Conjugate is dispensed onto the For In Vitro Diagnostic Use.
matrix cell and binds with any microparticle-bound antibody-antigen-
antibody complex.
The matrix cell is washed to remove materials not bound to the CAUTION: This product contains human sourced and/or
microparticles. potentially infectious components. For a specific listing, refer to the
The substrate, 4-Methylumbelliferyl Phosphate, is added. The alkaline Reagents section of this package insert. Components sourced from
phosphatase-labeled conjugate catalyzes the removal of a phosphate human blood present in the Positive Control have been tested and
group from the substrate, yielding the fluorescent product, found to be reactive for HBsAg and nonreactive for HIV-1 Ag and
4-Methylumbelliferone. This fluorescent product is measured by the MEIA antibodies to HCV, HBs and HIV-1/HIV-2. Components sourced from
optical assembly. human blood present in other kit components have been tested and
The presence or absence of HBsAg in the sample is determined by comparing found to be nonreactive for HBsAg and HIV-1 Ag and for antibodies to
the rate of formation of fluorescent product to a cutoff rate determined from HBs, HIV-1/HIV-2 and HCV. No known test method can offer complete
a previous AxSYM HBsAg (V2) Index Calibration. If the rate of formation of assurance that products derived from human sources or inactivated
fluorescent product in the test sample is greater than or equal to the cutoff microorganisms will not transmit infection. Therefore, all human
rate, the sample is considered reactive for HBsAg. sourced materials should be considered potentially infectious.
For further information regarding MEIA technology, refer to the AxSYM It is recommended that these reagents and human specimens be
System Operations Manual, Section 3. handled in accordance with the OSHA Standard on Bloodborne
Pathogens.18 Biosafety Level 219 or other appropriate biosafety
REAGENTS
practices20,21 should be used for materials that contain or are suspected
REAGENT PACK, 100 TESTS of containing infectious agents. These precautions include, but are
AxSYM HBsAg (V2) Reagent Pack (7A40-22) not limited to the following:
1 Bottle (5.1 mL) Anti-HBs (Mouse, Monoclonal, IgM) Coated 1. Wear gloves when handling specimens or reagents.
Microparticles in Phosphate buffer with protein stabilizers. Minimum 2. Do not pipette by mouth.
concentration: 0.15%. Preservative: Sodium Azide. (Reagent Bottle 2) 3. Do not eat, drink, smoke, apply cosmetics, or handle contact lenses in
1 Bottle (15.5 mL) Anti-Biotin (Rabbit):Alkaline Phosphatase Conjugate areas where these materials are handled.
in TRIS buffer with (Rabbit) IgG. Minimum concentration: 0.03 g/mL. 4. Clean and disinfect all spills of specimens or reagents using a
Preservative: Sodium Azide. (Reagent Bottle 1) tuberculocidal disinfectant such as 0.5% sodium hypochlorite, or other
suitable disinfectant.22,23

2
5. Decontaminate and dispose of all specimens, reagents and other INSTRUMENT PROCEDURE
potentially contaminated materials in accordance with local, state and Assay File Installation
federal regulation.24
The AxSYM HBsAg Assay File must be installed on the AxSYM System from
SAFETY PRECAUTIONS the software disk, 9A16-03, or higher, prior to performing the HBsAg (V2)
The AxSYM Probe Cleaning Solution (2% TEAH) may cause mild eye assay. Refer to the AxSYM System Operations Manual, Section 2, for proper
irritation. If this solution comes in contact with eyes, rinse immediately installation procedures.
with water. If irritation persists, seek medical attention. AxSYM HBsAg (V2) Assay Parameters
All components of this product contain Sodium Azide. For a specific listing
refer to the Reagents section of this package insert. The components The default assay parameters for the AxSYM HBsAg (V2) assay are listed
containing Sodium Azide are classified per the applicable European below. These parameters can be printed, displayed and edited according to
Community (EC) Directives as: Harmful (Xn). The following are the the procedure in the AxSYM System Operations Manual, Section 2. To print
appropriate risk (R) and safety (S) phrases. the assay parameters, press PRINT. Assay parameters that can be edited
contain a (>) symbol. In order to obtain values for the parameters with an
R22 Harmful if swallowed.
asterisk (*), review the specific Assay Parameter screen.
R32 Contact with acids liberates very toxic gas.
S35 This material and its container must be disposed of in Assay Parameters
a safe way.
1 Long Assay Name (English): HBsAg
S36 Wear suitable protective clothing.
6 Abbrev Assay Name (English): HBsAg
S46 If swallowed, seek medical advice immediately and
show this container or label. 11 Assay Number: 106
12 Assay Version: 2
Information for European customers: For product not classified as 13 Calibration Version: 00
dangerous per European Directive 1999/45/EC - Safety data sheet 14 Assay File Revision: *
available for professional user on request. 15 Assay Enabled > ON
17 Assay Type: MEIA
HANDLING PRECAUTIONS
18 Standard Cal Reps > 5
AxSYM HBsAg (V2) reagents are susceptible to splashes, air bubbles, 19 Master Cal Reps: 0
foaming and require inspection and removal of bubbles before loading. If 20 Cal Adjust Reps: 0
bubbles are present, refer to the AxSYM System Operations Manual, 21 Cal A Concentration: 1.00
Section 9: Service and Maintenance, Subsection: Daily Maintenance. 22 Cal B Concentration: 0.00
Do not use Solution 1 (MUP) beyond the expiration date or a 23 Cal C Concentration: 0.00
maximum of fourteen days on-board the AxSYM System. When 24 Cal D Concentration: 0.00
loading new Solution 1 (MUP), it is important to immediately tighten 25 Cal E Concentration: 0.00
the instrument cap for MUP to minimize exposure to air. Prolonged 26 Cal F Concentration: 0.00
exposure to air may compromise performance. 27 Master Calibrator 1 Concentration: 0.00
Do not use kits beyond the expiration date or a maximum of 112 cumulative 28 Master Calibrator 2 Concentration: 0.00
hours on-board the AxSYM System. 29 Cal Adjust Concentration: 0.00
Do not mix reagents from different reagent packs. Do not mix reagent 43 Default Dilution Protocol > UNDILUTED
packs and index calibrators from different lots. 44 Default Calibration Method > Index Cal
Avoid microbial contamination of samples and reagents. Use of disposable 45 Selected Result Concentration Units > S/N
pipettes or pipette tips is recommended. 46 Selected Result Decimal Places > 2
Avoid chemical contamination of reagents and equipment. 62 Blank I-Max background intensity: 0.0000
63 Min Tracer-Min net intensity: 0.0000
Ensure that sufficient sample volume is present. If sample volume is
64 Max Intercept-Max MUP intercept : 10000.0000
insufficient, the AxSYM System will give an error code and no result will
65 Min Intercept-Min MUP intercept : 1200.0000
be reported. For a description of the System error codes, refer to the
66 Upper limit for NRMSE for low rates : 9999.9900
AxSYM System Operations Manual, Section 10. 67 Upper limit for NRMSE for high rates : 0.5000
Inadequate adherence to package insert instructions may result in 68 Max Rate-Max rate used to check Min MUP Intercept : 50.0000
erroneous results. 69 Min Rate-Rate cutoff for NRMSE and Corr. Coef. : 8.0000
Use accurately calibrated equipment. 70 Min correlation coefficient for low rates : 0.9500
Use caution in handling patient samples to prevent cross contamination. 71 Min correlation coefficient for high rates : 0.9700
Transfer of as little as 1 L from one HBsAg positive sample may 72 MUP T Delay-Time delay following MUP: 2.6000
contaminate an adjacent negative sample and result in a falsely reactive 73 Low Limit-Normal/Therapeutic Range lower limit > 0.00
result. 74 High Limit-Normal/Therapeutic Range upper limit > 0.00
Additional safety and handling precautions and limitations for the reagent 75 Low Extreme Value > 0.00
packs, index calibrators, controls, patient samples and other reagents are 76 High Extreme Value > 0.00
described in the AxSYM System Operations Manual, Sections 7 and 8. 77 Lo Norm-% Uptake Normal Range Low > 0.0000
78 Hi Norm-% Uptake Normal Range High > 0.0000
STORAGE INSTRUCTIONS 80 Interpretation Option to use > 1
The AxSYM HBsAg (V2) Reagent Pack, Index Calibrator 84 Hold results with POS interpretation > ON
and AxSYM HBsAg Controls must be stored at 2-8C. The 85 Hold results with NEG interpretation > OFF
Reagent Pack, Index Calibrator and Controls may be used 86 Hold results with GRY interpretation > ON
91 Low Range Neat: 0.00
immediately after removal from the refrigerator. Do not
92 High Range Neat: 3000.00
freeze the reagents.
96 Low Range Dil1: 0.00
The AxSYM HBsAg (V2) Reagent Pack may be on-board the AxSYM System
97 High Range Dil1: 0.00
for a maximum of 112 cumulative hours. After 112 hours the reagent pack 101 Low Range Dil2: 0.00
and its associated index calibrator must be discarded. Refer to the AxSYM 102 High Range Dil2: 0.00
System Operations Manual, Appendices, for further information on tracking 106 Low Range Dil3: 0.00
on-board time. 107 High Range Dil3: 0.00
Solution 1 (MUP) must be stored at 2-8C. It may be on-board the AxSYM 117 Negative Interpretation Cutoff > 2.00
System for a maximum of fourteen days. After fourteen days it must be
discarded. NOTE: Parameters 18, 43, 44 cannot be edited to another value.
The AxSYM Probe Cleaning Solution, Solution 3 (Matrix NOTE: Parameter 45 can be edited to the alternate Result Unit S/CO.
Cell Wash) and Solution 4 (Line Diluent) must be stored at Refer to the AxSYM System Operations Manual for a detailed description of
15-30C. Instrument Procedures.
INDICATIONS OF INSTABILITY OR DETERIORATION OF NOTE: A grayzone feature is available. To utilize the grayzone option,
REAGENTS parameter 117, Negative Interpretation Cutoff, must be edited to desired
value within the range of 1.60 to 1.99 S/N or 0.80 to 0.99 S/CO. Specimens
When an AxSYM HBsAg Positive or Negative Control value is out of the
with S/N values equal to the selected value and less than 2.00 or with S/CO
expected range, it may indicate deterioration of the reagents or errors in
values equal to the selected value and less than 1.00 are indicated as
technique. Associated test results are invalid and must be retested. Assay
"GZ-NEGATIVE" or "GZ-NONREACTIVE" in the interpretation field. The
recalibration may be necessary.
report options are selected in parameter 80, Interpretation Option to use.

3
The AxSYM HBsAg (V2) values available for parameter #80 (Interpretation
Option to use) are:
POS Interp NEG Interp GRY Interp
1. REACTIVE NEGATIVE GZ-NEGATIVE
2. REACTIVE NONREACTIVE GZ-NONREACTIVE
3. REACTIVE BLANK BLANK
NOTE: Parameter 46 must be set to 3 in order to edit the alternate Result
Unit S/CO grayzone value to 0.80.
SAMPLE COLLECTION AND PREPARATION FOR ANALYSIS
Serum (including serum collected in separator tubes) or plasma (collected
in sodium heparin, sodium citrate, ACD, CPDA-1 or EDTA) may be used
in the AxSYM HBsAg (V2) Assay. Use the ratio of anticoagulant (quantity)
to sample (volume) that is recommended by the manufacturer.
The AxSYM System does not have the capability to verify the sample
type. It is the responsibility of the operator to verify the correct sample
type(s) are used in the AxSYM HBsAg (V2) assay.
Samples containing particulate matter or red blood cells may give
inconsistent or erroneous results.
Inspect all samples for splashing, air bubbles and foaming. If necessary
remove bubbles prior to analysis using a new applicator stick for each
sample. Refer to the AxSYM System Operations Manual, Section 7.
Specimens with obvious microbial contamination should not be used.
All patient samples to be tested in Primary Tubes must be centrifuged
to remove red cells or particulate matter. Each sample that requires
repeat or confirmatory testing, or that has been frozen and thawed,
must be transferred to a centrifuge tube and centrifuged at an RCF*
of at least 10,000 x g for 10 minutes. Transfer clarified sample to a
sample cup for testing.
WARNING: AxSYM System Software Version 3.0 and higher offers an
Auto Retest/Auto Dilution feature. Due to requirements discussed above,
this feature must not be used.
Samples which contain fibrin may cause erroneous results. Serum
samples must be completely clotted and centrifuged prior to testing to
generate consistent results.
Centrifuged specimens with a lipid layer on top of the liquid must be
transferred to a sample cup or secondary tube. Care must be taken to
transfer only the clarified specimen and not the lipemic material.
Samples from heparinized patients may be partially coagulated and
erroneous results could occur due to the presence of fibrin. To prevent
this phenomenon, draw the sample prior to heparin therapy.
Samples may be stored on the clot or red blood cells for up to 14 days at
2-8C prior to being tested.
If the assay will be performed after 14 days, the sample must be removed
from the clot or red blood cells and stored frozen (-10C or colder).
15 nonreactive and 15 reactive samples showed no qualitative
performance differences when subjected to 6 freeze-thaw cycles; however,
multiple freeze-thaw cycles should be avoided. Samples must be mixed
thoroughly after thawing and centrifuged prior to use, as described in this
section, to remove particulate matter and to ensure consistency in the
results.
When shipped, samples must be packaged and labeled in compliance
with applicable federal and international regulations covering the transport
of clinical samples and etiologic agents. Samples may be shipped under
ambient conditions, refrigerated on wet ice (2-8C) or frozen on dry ice
(-10C or colder).
All samples (patient samples, controls and calibrators) should be tested
within 3 hours of being placed on-board the AxSYM System. Refer to the
AxSYM System Operations Manual, Section 5, for more detailed
discussion of on-board sample storage constraints.
Do not use heat-inactivated samples.
Performance has not been established for cadaver samples or body fluids
such as urine, saliva, semen or amniotic fluid.
No qualitative performance differences were observed when HBsAg
Negative and Positive Controls were tested with elevated levels of bilirubin
(0-20 mg/dL), hemoglobin (0-250 mg/dL) or lipids (0-1000 mg/dL).
The assay was designed and validated for use with human serum or
plasma from individual patient and donor specimens. Pooled specimens
should not be used since the accuracy of their test results has not been
validated.
* RCF = Relative Centrifugal Force
4
SAMPLE VOLUME
The sample volume required to perform a single HBsAg (V2) test on the
AxSYM System varies according to the different sample containers. For
sample cups, the minimum sample volume required is 190 L. For every
additional HBsAg (V2) test performed from the same sample container, an
additional 140 L of sample will be required.
The sample cup minimum volume will be calculated by the AxSYM System.
It will be displayed on the Order screen at the time the test(s) is(are) ordered
and printed in the Orderlist Report.
To obtain the recommended volume requirements for the AxSYM HBsAg
(V2) Index Calibrator and AxSYM HBsAg Controls, hold the bottles vertically
and dispense 15 drops of Index Calibrator or 5 drops of Positive or Negative
Control into each respective sample cup.
For sample volume requirements in primary or aliquot tubes, and control
volume requirements for multiple AxSYM HBsAg (V2) reagent lots, refer to
the AxSYM System Operations Manual, Section 5.
AxSYM HBsAg (V2) PROCEDURE
Materials Required
7A40-22 AxSYM HBsAg (V2) Reagent Kit, containing:
AxSYM HBsAg (V2)
AxSYM HBsAg (V2)
100
100
7A40-10 AxSYM HBsAg Controls
8A47-04
8A81-04
8A46-01
9A35-04/ AxSYM
9A35-05
8A76-01
Materials Required But Not Provided
Pipettes or pipette tips
CAUTION:
When manually dispensing samples, verify that dispensing equipment
does not introduce cross contamination and that it delivers the specified
sample volume. Use a separate disposable pipette or pipette tip for each
sample.
For optimal performance, it is important to follow the routine maintenance
procedures defined in the AxSYM System Operations Manual, Section
9. If your laboratory requires more frequent maintenance, follow those
procedures.
Assay Procedure
CAUTION: The System status must be WARMING, PAUSED, READY or
STOPPED before adding or removing sample segments, reagent packs or
reaction vessels.
1. Check for sufficient on-board inventory of matrix cells and bulk solutions,
and for sample segment availability.
2. Check for sufficient waste collection capacity.
CAUTION: Do not open the Interior Waste Door or the AxSYM Processing
Center Cover while any test is in progress. If opened, all processing will
stop. All tests will be terminated and must be repeated.
3. Order the HBsAg (V2) Index Calibrator, HBsAg Controls, and/or patient
samples as required. Assign or modify sample segment position (S/P)
for each sample, as necessary.
Refer to the Quality Control Procedures section in this package insert for
calibration and control requirements.
Index Calibration:
Perform AxSYM HBsAg (V2) calibration by testing 5 replicates of the
Index Calibrator. Dispense 15 drops of the Index Calibrator into a sample
cup. Do not simultaneously calibrate more than one AxSYM HBsAg (V2)
reagent lot.
Controls:
Perform quality control by testing the Positive and the Negative Control
(one test each). Dispense 5* drops each of Positive and Negative Controls
into individual sample cups.
* When more than one AxSYM HBsAg (V2) reagent lot is on-board the AxSYM
System, multiply control volume by the number of lots.
 Patient Samples:
Ensure that sufficient volume is present in the sample cups or tubes. The
sample cup minimum volume is 190 L for the first HBsAg (V2) test plus
140 L for each additional HBsAg (V2) test. For volume requirements in
primary or aliquot tubes, refer to the AxSYM System Operations Manual,
Section 5.
NOTE: The operator may obtain an Orderlist Report by pressing PRINT.
The printout contains sample placement information and minimum sample
cup volume requirements for all tests ordered. When using Host Order
Query, the Order screen information and the Orderlist Report is not
available. Refer to the AxSYM System Operations Manual, Section 5:
Ordering Patient Samples, for a description of the Host Query Option.
4. Place sample segments containing the ordered samples into the Sample
Carousel.
5. Place the HBsAg (V2) Reagent Pack into the Reagent Pack Carousel.
6. Ensure that reaction vessels (RVs) are present on the RV Carousel.
Additional RVs may be added as needed.
7. Press RUN. All entries on the Orderlist screen are transferred to the Order
Status screen for sample processing.
8. Review the results to determine whether retesting is required.
9. When testing is completed, remove the samples and the AxSYM HBsAg
(V2) Reagent Pack from the Sampling Center. Store at 2-8C.
Refer to Sections 5 and 6 of the AxSYM System Operations Manual for a
detailed explanation of performing assay calibration and sample testing
procedure.
QUALITY CONTROL PROCEDURES
CALIBRATION
Five replicates of the HBsAg (V2) Index Calibrator must be tested for an
AxSYM HBsAg (V2) Calibration. A single sample of both the Positive and
Negative Controls must be tested as a means of evaluating the assay
calibration.
Once an AxSYM HBsAg (V2) calibration is accepted and stored, all
subsequent samples may be tested without further calibration unless:
A reagent pack with a new lot number is used for an assay.
Either of the AxSYM HBsAg Control values is out of its specified range.
Refer to the AxSYM System Operations Manual, Section 6, for additional
information.
The operator must verify that the AxSYM HBsAg Control values are within
the acceptable ranges specified in this package insert (see Controls section).
The AxSYM System verifies that the results of an assay calibration meet the
specifications assigned to selected validity parameters. An error message
occurs when the calibration fails to meet a specification. Refer to the AxSYM
System Operations Manual, Section 10, for an explanation of the corrective
actions for the error code. Refer to the AxSYM System Operations Manual,
Appendix E, for an explanation of the calibration validity parameters that
may be used by the AxSYM System.
QUALITY CONTROL
The minimum control requirement for an AxSYM HBsAg (V2) assay is a
single sample of each of the Positive and Negative Controls tested once
every 24 hours, each day of use. Controls may be placed in any position in
the Sample Carousel.
The AxSYM HBsAg Control values must be within the acceptable ranges
specified in this package insert (see Controls section). If a control value is
out of its specified range, the associated test results are invalid and must be
retested. Recalibration may be indicated.
If the quality control procedures in your laboratory require more frequent
use of controls, follow those procedures.
Fluorescence Background Acceptance Criteria
Quality Control with regards to the MUP substrate blank is automatically
determined by the instrument and checked under Assay Parameter 64 Max
Intercept-Max MUP intercept each time a test result is calculated. If the MUP
intercept value is greater than the maximum allowable value, the result is
invalid. The test request will be moved to the Exceptions List where it will
appear with the message "1064 Invalid test result, intercept too high" and
the calculated intercept value. Refer to the AxSYM System Operations
Manual, Section 10, when this error message is obtained.
Refer to the AxSYM System Operations Manual, Section 2, for further
information on parameter files.
RESULTS
CALCULATIONS
The AxSYM System calculates the Cutoff rate from the mean rate of five
Index Calibrator replicates and stores the result. The AxSYM HBsAg (V2)
assay has two selectable Result Units. One reports the result as a ratio S/N
and the other reports the result as a ratio S/CO. These units can be selected
in the assay parameters according to the procedure in the AxSYM System
Operations Manual, Section 2.
5
The Cutoff rate is determined by multiplying the Index Calibrator mean rate
by 2.
Cutoff Rate (CO) = Index Calibrator mean rate x 2
S/N
The AxSYM HBsAg (V2) assay calculates a result based on the ratio of the
sample rate to the stored Index Calibrator mean rate for each sample and
control.
Sample Rate
S/N =
Index Calibrator Mean Rate
S/CO
The AxSYM HBsAg (V2) assay calculates a result based on the ratio of the
sample rate to the Cutoff rate (CO) for each sample and control.
Sample Rate
S/CO =
Cutoff Rate (CO)
FLAGS
Some results may contain information in the Flags field. For a description of
the flags that may appear in this field, refer to the AxSYM System Operations
Manual, Section 1 and 2.
INTERPRETATION OF RESULTS
Samples with S/N less than 2.00 are negative by the AxSYM HBsAg (V2)
assay and need not be tested further.
Samples with S/N values greater than or equal to 2.00 are considered
reactive.
Samples with S/CO values less than 1.00 are negative by the AxSYM
HBsAg (V2) assay and need not be tested further.
Samples with S/CO values greater than or equal to 1.00 are considered
reactive.
All samples that are reactive on initial testing should be retested in
duplicate using the AxSYM HBsAg (V2) assay. If neither of the retests
are reactive, the sample must be considered negative for HBsAg. If the
sample is reactive in either of the repeated replicates, the sample must
be considered repeatedly reactive.
Repeatedly reactive samples should be tested by a neutralizing
confirmatory test, such as the AxSYM HBsAg Confirmatory assay.
Samples which are confirmed by neutralization with human anti-HBs must
be considered positive for HBsAg.
False reactive results may be obtained with any diagnostic test. Two types
of false reactive results may occur with AxSYM HBsAg (V2), nonrepeatable
reactives and nonspecific reactives.
Nonrepeatable Reactives: Some samples which are reactive in the AxSYM
HBsAg (V2) assay may not be reactive on retesting. The most common
sources of nonrepeatable reactives are:
particulate matter in the patient sample, particularly fibrin clots and
cellular material.
contamination of nonreactive samples caused by transfer of a high
titer antigen sample.25
Nonspecific Reactives: All highly sensitive immunoassay systems have a
potential for nonspecific reactions. The specificity of a repeatedly reactive
sample should be confirmed by a neutralizing confirmatory test, such as the
AxSYM HBsAg Confirmatory assay. A nonspecific reactive sample will be
repeatedly reactive but will not be confirmed by neutralization. Therefore, it
is recommended that a specific antibody neutralization be performed prior
to informing the donor/patient of their HBsAg carrier status. For additional
information on neutralization testing, refer to the AxSYM HBsAg Confirmatory
assay package insert.
LIMITATIONS OF THE PROCEDURE
For diagnostic purposes and in order to differentiate acute HBV infection
from chronic HBV infection, the detection of HBsAg must be correlated
with patient symptoms and other hepatitis B viral serological markers.
It is recognized that presently available methods for the detection of
hepatitis B surface antigen may not detect all potentially infectious units
of blood or infected individuals. A nonreactive test result does not exclude
the possibility of exposure to or infection with hepatitis B. Nonreactive
test results in individuals with prior exposure to hepatitis B may be due to
antigen levels below the detection limit of this assay or lack of antigen
reactivity to the antibodies used in this assay.
This assay was designed and validated for use with human plasma or
serum from individual patient and donor specimens. Insufficient data are
available to interpret tests performed on pooled blood or processed plasma
and products made from such pools. Testing of these specimens is not
recommended.
Performance has not been established using cadaver samples or body
fluids such as urine, saliva, semen or amniotic fluid.
 Do not use heat-inactivated samples. TABLE 3
Frozen samples and those containing particulate matter or red blood Number (%) of Reactive Samples Detected by AxSYM HBsAg (V2) and
cells must be centrifuged prior to running the assay. Confirmed as Positive for HBsAg
Samples which contain fibrin may cause erroneous results. Serum Number of Initially Repeatedly
samples must be completely clotted to generate consistent results. samples reactive reactive Confirmed
Samples from heparinized patients may be partially coagulated. Erroneous Group tested (%) (%) Positivea Specificity
results could occur due to the presence of fibrin. To prevent this
phenomenon, draw the sample prior to heparin therapy. Blood Donors 2003 2 (0.10) 1 (0.05) 0 99.95 %
Blood Donors 2032 1 (0.05) 1 (0.05) 0 99.95 %
EXPECTED VALUES (including 401
In random blood donor populations, the number of samples found repeatedly first donors)
reactive for HBsAg by AxSYM HBsAg (V2) has typically been less than
Hospital Patients 503 2 (0.40) 2 (0.40) 0 99.60 %
0.20%. In a population reactive for HBsAg, AxSYM HBsAg (V2) detected
HBsAg in 402/402 (100%) specimens. Medical Conditions 50 2 (4.00) 0 (0) 0 100.00 %
and Potentially 57 1 (1.75) 1 (1.75) 1 100.00 %
SPECIFIC PERFORMANCE CHARACTERISTICS Interfering
PRECISION Substances
Assay reproducibility was determined during the clinical evaluation of AxSYM (both unrelated to
HBsAg. The six member panel was run in replicates of three, four times per HBV Infection)b
day, over two days on each of three clinical lots. Calculations were made a A confirmed positive result in these studies was defined as neutralization by
with a variance components analysis, using a nested analysis of variance AxSYM HBsAg Confirmatory Assay.
model.26 The results from these three clinical lots are summarized in Table 1. b Medical Conditions Unrelated to HBV Infection and Potentially Interfering

TABLE 1 Substances included the following categories of previously frozen specimens:

high titre Immunoglobulin (IgG Myeloma, IgM Myeloma, Pregnant Females 1st,
Reproducibility of AxSYM HBsAg
2nd and 3rd trimester, Flu Vaccine Recipients), antibodies to E. coli, anti-CMV
Panel Mean Intra-Run Inter-Run Total positive, anti-EBV positive, anti-HSV positive, rubella antibody positive, yeast
Member N S/N SD %CV SD %CV SD %CV infection, anti-nuclear antibody positive, syphilis positive, rheumatoid factor and
toxoplasmosis antibody positive.
1 72 4.12 0.13 3.1 0.13 3.3 0.36 8.7
2 72 2.67 0.11 4.0 0.11 4.1 0.24 8.9 The specificity of AxSYM HBsAg (V2) was 99.95% (4033/4035) in volunteer
3 72 1.57 0.07 4.2 0.07 4.6 0.12 7.7 blood donors based on an assumed zero prevalence of HBsAg. In this
4 72 5.63 0.18 3.2 0.19 3.3 0.35 6.3 calculation no repeatedly reactive volunteer blood donor sample was excluded
5 72 3.63 0.13 3.6 0.13 3.7 0.25 6.7
due to confirmation by neutralization with anti-HBs.
6 72 1.84 0.08 4.3 0.08 4.5 0.13 7.0
The specificity of AxSYM HBsAg (V2) was 99.60% (501/503) in hospital
patients. Two of 503 specimens obtained from hospital patients were repeatedly
Index 48 0.96 0.06 5.8 0.06 5.8 0.06 6.3
reactive. In this calculation no repeatedly reactive sample was excluded due
NC 48 0.98 0.06 5.7 0.06 5.7 0.06 6.2
to confirmation by neutralization with anti-HBs.
PC 48 21.24 0.80 3.8 0.82 3.9 2.32 10.9
In 107 specimens from indiviuals with medical conditions unrelated to HBV
Index = Index Calibrator infection and specimens containing potentially interfering substances, one
NC = Negative Control specimen was repeatedly reactive and confirmed positive for HBsAg.
PC = Positive Control
SENSITIVITY
DETECTABILITY
The sensitivity of AxSYM HBsAg (V2) was evaluated using a six member
The ability of AxSYM HBsAg (V2) to detect HBsAg in specimens from blood proficiency panel of purified HBsAg (ad and ay) prepared at Abbott Laboratories.
donors, hospital patients (samples submitted to hospital laboratories for Each panel member was assayed in replicates of 3 on each of two instruments.
diagnostic testing) and hepatitis B patients is shown in Table 2. The data Typical results are summarized in Tables 4, 5 and 6.
includes a total of 4940 serum and plasma specimens.
TABLE 4
TABLE 2 Detection of Purified HBsAg/ad
Detection of HBsAg in Serum and Plasma Specimens from Blood
Donors, Hospital Patients and Hepatitis B Patients Concentration AxSYM HBsAg (V2)
(ng/mL) S/N Value Result
AxSYM HBsAg (V2)
Population Number Confirmed 0.721 4.35 +
Group Tested Number (%) Positive (%) 0.496 3.57 +
0.229 2.26 +
Blood Donors 4035 0 (0) 0 (0)
Hospital Patients 503 0 (0) 0 (0) TABLE 5
Hepatitis B Patients 402 402 (100) 402 (100) Detection of Purified HBsAg/ay
SPECIFICITY Concentration AxSYM HBsAg (V2)
The percentage of samples found to be initially reactive with AxSYM HBsAg (V2) (ng/mL) S/N Value Result
and the percentage of these samples found to be repeatedly reactive were 0.722 6.09 +
determined by testing 4538 samples (plasma and sera) obtained from two 0.498 4.59 +
blood banks and a hospital laboratory in a clinical evaluation. The presence 0.250 2.88 +
of HBsAg in the repeatedly reactive specimens was confirmed by neutralization
with anti-HBs using AxSYM HBsAg Confirmatory assay. The results of these TABLE 6
tests are shown in Table 3. Sensitivity Calculated by Linear Regression Analysis
HBsAg Sensitivity
Subtype (ng/mL)
ad 0.19
ay 0.14
Typically AxSYM HBsAg (V2) sensitivity results from clinical trials and studies
done at Abbott Laboratories have ranged from 0.10 to 0.60 ng/mL.
In studies performed at Abbott Laboratories using HBsAg ad reference serum
from Paul Ehrlich Institute (PEI), the AxSYM HBsAg (V2) assay calculated
sensitivity was 0.031 PEI Units/mL.

6
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