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US Army Hospital Heidelberg, Germany
Pro 68-0.1
9 June 2008
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Heidi
SMILE Comments: This document is provided as an example only. It must be revised to accurately reflect
your labs specific processes and/or specific protocol requirements. Users are directed to countercheck
facts when considering their use in other applications. If you have any questions contact SMILE.
Copy # _____
U. URINALYSIS
U.02. MANUAL URINALYSIS MICROSCOPIC EXAM
U.2.1.
PRINCIPLE:
1. Routine Urinalysis consists of both physical and chemical analyses to assist physicians
in the diagnosis and treatment of renal and urinary tract diseases and in the detection
of metabolic or systemic disease processes not directly related to the kidney.
2. The microscopic examination of the centrifuged urine sediment includes the study of
formed elements, such as WBCs, RBCs, casts and crystals.
3. The macroscopic examination of urine includes physical appearance, such as color,
character and clarity. See U.1 Manual Urinalysis Dipstick SOP for macroscopic
testing and reporting procedures.
4. A qualitative chemical analysis of the urine is performed by using a multi-parameter
test strip that measure pH, protein, glucose, ketones, bilirubin, urobilinogen, nitrite,
blood, leukocyte esterase, and specific gravity. The test strips are dipped in the urine
and read visually according to the color comparison chart printed on the side of the
container at prescribed time intervals. See U.1 Manual Urinalysis Dipstick SOP for
qualitative testing and reporting procedures.
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to 8C. (The specimen must be brought to room temperature before testing.)
5. The following urine samples are not satisfactory for testing:
5.1. Specimens received over two hours after collection.
5.2. Mislabeled samples.
5.3. Improperly collected samples. For example, urine samples with preservatives,
specimens collected in non-sterile containers, or specimens collected in
containers with soap or detergent residues will not be accepted.
5.4. QNS (Quantity Not Sufficient) - The recommended minimum volume is 12 mLs.
The required minimum volume for microscopic examination is 0.50 mL. In the
event that less than <0.50 mL is received, perform testing on uncentrifuged urine
and comment as noted in section U.2.7.
5.5. In the event that an unacceptable sample is received, another sample must be
requested. Document all action taken in the LIS.
U.2.4. EQUIPMENT & REAGENTS
1. Equipment:
1.1
1.2.
1.3.
1.4.
1.5.
1.6.
1.7.
2. Reagents - None
U.2.5. CALIBRATION: Not applicable
U.2.6. QUALITY CONTROL:
1. Control Materials and Procedural Notes:
1.1. A Run in urinalysis is defined as all patient testing performed in a 24-hour period.
Quality Control must be performed under the following conditions.
1.1.1. Every morning prior to testing patient specimens.
1.1.2. Whenever test results appear questionable.
1.2. A Normal and Abnormal Kova Control will be performed at least once every
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run.(see above)
1.3. Reconstitute new Kova controls with 60 mL Reagent Grade water according
to manufacturers instructions.
1.3.1. After reconstitution, separate controls into 2 mL aliquots using
disposable transfer pipettes and plastic test tubes. Cap tightly!
1.3.2. Label each plastic test tube with Normal or Abnormal Kova Control,
lot number, expiration date, reconstitution date, and initials of tech who
reconstitutes controls.
1.3.3. Sample aliquots will be stored frozen at -20C. The sample aliquots are
stable at this temperature for up to four (4) months.
1.3.4. Only one freeze/thaw cycle is permitted. Aliquots must be at room
temperature before testing. Do not use a warming block to thaw.
1.3.5. Sample aliquots are stable for one hour at 15-20C or 24 hours at 2-8C.
1.4. Perform the Kova controls in the same manner as patient samples. See section
U.2.7 for the patient testing procedure. Record the results on the Urinalysis
Quality Control Worksheets. See appendix 3 & 4.
1.5. All results must fall into the expected ranges, which are obtained from the
manufacturers insert for each control and recorded on the top of the Urinalysis
Quality Control Worksheets.
1.6. If all results are within expected ranges, proceed with patient testing.
1.7. If QC results are outside of expected ranges for controls:
1.7.1. Re-examine the specimen using a different field of view.
1.7.2. Adjust the coarse/fine focus as needed to improve the view and repeat
testing.
1.7.3. Check for signs of contamination in the controls.
1.7.4. Repeat the procedure with freshly reconstituted Kova controls.
1.7.5. If the results are still out of range after performing the above steps, notify
the next higher supervisor immediately. Corrective action must be taken
and QC must be in range before patient testing can be performed.
1.7.7. Record all QC values (both in and out of range) and corrective actions
on the Quality Control Worksheets, appendix 3 & 4.
1.8. Parallel Testing: All new quality control lots or separate shipments of current
lots will be tested against the current lots for performance verification before
performing patient testing. See the Parallel Testing form, Appendix 5. Results
are considered acceptable if they are within their lot number quality control
limits.
U.2.7. PROCEDURE
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1. Remove the specimens/controls from the refrigerator or freezer, as applicable, and
allow them come to room temperature for at least 30 minutes before mixing.
2. Positively identify the specimens using the patient name and date of birth.
3. Log the sample on the testing worksheet using the LIS generated patient label,
appendix 6.
4. Add the LIS generated label to the sample cup (not the sample lid). Any discrepancy
must be investigated before processing the specimen.
5. Mix patient urine sample by swirling. Mix control aliquots by inverting several times
to ensure homogeneity of the contents.
6. If a macroscopic examination & urine chemistry testing is ordered refer to U.01.1
Manual Urinalysis Dipstick Testing SOP for testing and reporting procedures. The
microscopic examination is performed after dipstick testing unless the provider
requests otherwise.
7. If Clintest, Acetest or Ictotest is required as listed below or requested by the provider,
perform testing on uncentrifuged urine prior to performing the microscopic exam.
Refer to individual confirmatory test SOPs as listed in the references.
8. The Clinitest confirmatory test is required for all pediatric urine samples with
patients age 2 years old or younger, regardless of whether it was requested or not.
See Clinitest SOP U06.1.
9. Backup tests and/or microscopic examination must be performed on urine that
tests positive for the following:
Positive Result
Backup Test
Microscopic Exam
Blood
N/A
Yes
Nitrite
N/A
Yes
Ketones
Acetest
No
Glucose
Clinitest
No
Bilirubin
Ictotest
No
Protein
3% Sulfosalicylic Acid
Yes
Leukocyte Esterase
N/A
Yes
10. Confirmatory tests are not required for TRACE results on PROTEIN or KETONES.
11. Microscopic examinations are required on the following specimens regardless of the
reagent strip results:
11.1. Urine with cloudy or hazy appearance.
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11.2.
11.3.
11.4.
11.5.
12. Pour 12-mLs of specimen into a sterile Kova tube and label the tube with the LIS
specimen label. If the total specimen volume is 0.50 mL or less, do not pour the
sample into a Kova tube. Perform testing from the specimen cup as indicated in the
following steps.
13. Testing
13.1. Volumes greater than 0.50 mL
13.1.1. Perform Dipstick, Clinitest, Acetest, or Ictotest as required or
requested.
13.1.2. Cap the Kova sample and centrifuge at 1500 rpm for 5 minutes.
13.1.3. Remove the sample from the centrifuge and discard the cap.
13.1.4. If 3% SSA testing is required or requested, perform testing on the
supernatant according to the U.10.1. 3% SSA SOP.
13.1.5. Decant and discard the supernatant while retaining the sediment pellet
in the conical portion of the Kova tube. There will be a small volume
of fluid retained above the sediment. The total volume retained in the
Kova system tube after decanting the supernatant should be
approximately 0.5 mL.
13.1.6. Use a Kova disposable pipette to resuspend the sediment pellet.
13.1.7. Proceed with testing in step 13.3.
13.2. Volumes less than 0.50 mL - swirl the sample in the specimen cup to mix well.
13.3. Use a Kova pipette to aspirate sample from the well mixed specimen.
13.4. Place one (1) drop of the sediment on the Kova slide so that the urine sediment
is drawn under the preformed Kova cover slip.
13.5. Place the Kova slide under the microscope with the low power objective (10X
lens) and observe the sediment for the presence of casts. Observe at least ten
(10) fields and take the average number of casts seen for each type.
13.6. Switch to high power objective (40X lens) to observe the sediment for the
presence of epithelial cells, WBCs, RBCs, bacteria, crystals, yeast, trichomonas,
mucus, and spermatozoa.
13.7. Scan approximately 8-10 fields and take the average per field for each type of
sediment.
13.8. QNS specimens - report the following canned comments as applicable:
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13.8.1. If the volume was <12 mL but > 0.5 mL, enter [12ML] Minimum specimen volume is 12 mL. Urine sediment results
may be falsely decreased due to a submitted sample volume less
than 12 mLs.
13.8.2. If the volume was <0.5 mL, enter [UNSPUN] - Testing performed
on uncentrifuged urine due to a submitted volume less than 0.5
mL. Minimum required specimen volume is 12 mL. Urine
sediment results may be falsely decreased.
13.9. Review the results and enter data on the manual microscopic patient result form
(appendix IV). See section U.2.8 for Expected and Critical Results - Abnormal
results will be flagged.
13.10.
Enter all results in the LIS. Do not certify any results until all urinalysis
testing ordered on the patient sample is complete.
13.11.Urine samples are retained at 2-10C for 24 hours in the Hematology
refrigerator.
U.2.8. EXPECTED / CRITICAL RESULTS & REPORTABLE RANGE:
1. The following bolded analytes require a result entry during the microscopic results
even if they are not seen during the microscopic exam. Report none or 0-1 or 0-2 as
indicated. All other analytes are optional entry if seen during the microscopic exam.
Analyte
HYALINE CASTS
0-1
Granular Cast
None
None
Waxy Cast
None
EPITHELIAL CELLS
0-2
LEUKOCYTES (WHITE
BLOOD CELLS)
0-2
0-2
BACTERIA
None - Few
MUCUS
None - Light
Crystals
None
Spermatozoa
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Reportable Range:
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U.2. Manual Urinalysis Microscopic Exam
Yeast
None
Trichomonas
None
2. If budding yeast or hyphea are present, add the following canned comments:
2.1. [Budding] - Budding yeast present.
2.2. [Hyphea] - Hyphae present.
3. The presence of sperm in the urine of a female child under 16 years old is a critical
value. Supervisor confirmation is required before reporting any suspected sperm to
the physician. The physician must be notified in accordance with critical value
procedures before sperm is reported in the LIS. The urine must be retained at 2-10C
until the patients physician and laboratory director authorizes disposal. In no instance
will the urine be destroyed until duly authorized.
4. If a urine dipstick is performed, compare the results obtained on the microscopic with
the multi-parameter reagent strip with the following facts in mind:
4.1.
Urine with RBCs seen on the microscopic exam should have a positive occult
blood on the reagent strip.
4.2.
Urine with casts should have elevated specific gravity and positive protein.
4.3.
4.4.
5. CRITICAL VALUES:
5.1. When the following values are encountered after a urinalysis testing,
immediately notify the physician or senior ward/clinic nurse of the values
encountered.
Analyte
Waxy Casts
Any
Any
Cystine Crystals
Any
Tyrosine Crystals
Any
Leucine Crystals
Any
Sperm
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5.2.
Critical values must be reported in accordance with the laboratory critical value
policy.
5.2.1. All critical microscopic sediment must be verified by the laboratory
supervisor before reporting to the physician/nurse and before certifying
results in the LIS.
5.2.2. All results must be read back by the physician/nurse and the
notification & read back documented in the LIS.
5.2.3. The pathologist must also be notified within 2 hours and will
determine if a pathologist review is required.
Alkaline Urine
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Abnormal
Acidic Urine
Alkaline Urine
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U.2. Manual Urinalysis Microscopic Exam
Amorphous Urates
Amorphous
Phosphates
Cystine
None
Uric Acid
Triple Phosphates
Tyrosine
Calcium Oxalate
Ammonium Biurates
Leucine
Calcium Phosphates
Sulfonamide
(Sulfadiazine)
Calcium Carbonates
2.7. Mucus: Light mucus is normally found in urine and can sometimes be confused
with hyaline casts.
2.8. Spermatozoa: A few are normally found in urine from normal males. Increased
numbers are found in prostatic disease.
2.9. Trichomonas: Normally not found in urine.
abnormal and may indicate infection.
Presence of trichomonas is
5.10. Yeast: 0-1/HPF or few per high power field are normal in females. Increased
numbers indicates infection.
5.11. Casts are classified according to their matrix, inclusions, pigments and cells
present.
5.11.1. Cast matrices:
5.11.1.1. Hyaline casts: Are translucent cylindrical structures composed
of mucoprotein. Increased numbers are seen with renal diseases
and transiently with exercise, fever, congestive heart failure, and
diuretic therapy.
5.11.1.2. Waxy casts: These differ from hyaline casts in that they are
easily visualized because of their high refractive index. Waxy
casts are homogeneously smooth in appearance. Their margins
are sharp, their ends are blunt, and cracks or convolutions are
frequently seen along the lateral margins. Waxy casts are
commonly associated with tubular inflammation and chronic
renal failure. They are also found during acute or chronic renal
allograft rejection.
5.11.2. Cast Inclusions:
5.11.2.1. Granular casts are semitransparent cylinders containing small
(fine) or large (coarse) granules. These granules represent
plasma protein aggregates.
Granular casts appear with
glomerular or tubular diseases.
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5.11.2.2. Fatty casts: Are semitransparent or granular cylinders
containing large highly refractive vacuoles or droplets. Visible
fat droplets are triglycerides or cholesterol esters. These are
commonly seen when there is heavy proteinuria and are a
feature of the nephrotic syndrome.
5.11.2.3. Crystal Casts: Crystalline inclusion in a semitransparent or
granular cylinder. These casts indicate disposition of crystals in
the tubule or collecting duct.
5.11.3. Cast Pigments:
5.11.3.1. Hemoglobin (Blood) casts: Appears yellow to red; sometimes
the color is very pale and difficult to interpret. These casts are
associated with glomerular disease.
5.11.3.2. Myoglobin casts: These cast are red-brown in color and occur
with myoglobinuria following acute muscle damage.
5.11.3.3. Bilirubin and other drug casts: Bilirubin is seen in urine when
there is obstructive jaundice, and will color casts as deep yellow
brown. Drugs such as phenazopyridine (Pyridium) cause a
bright yellow to orange color in acid urine and will color casts
and cells.
5.11.4. Cellular Casts:
5.11.4.1. Erythrocyte (Red Blood Cell) casts: Semitransparent or
granular cylinders containing distinct erythrocyte. Disorders
reflected in the presence of erythrocyte casts in the sediment
may include acute glomerulonephritis, IgA nephropathy, lupus
nephritis, subacute bacterial endocarditis, and renal infarction.
5.11.4.2. Leukocyte (White Blood Cell) Casts: Semitransparent or
granular cylinders containing leukocytes. They may be seen
pyelonephritis, glomerular diseases, interstitial nephritis, lupus
nephritis, and nephrotic syndrome.
6. SOURCES OF ERROR:
6.1. Urine should be tested within two (2) hours after collection. Prolonged testing
delay may result in cast dissolution, RBC crenation or bursting, increased
bacteria, and crystals dissolution.
6.2. Fill the Kova slide chamber with the pipette placed parallel to the slide and
dispense the specimen drop in the corner of the well. Samples placed in the
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center of the well can cause air bubbles to form resulting in sample distortion
during examination.
U.2.10. APPENDICES:
1.
2.
3.
4.
5.
6.
7.
U.2.11. REFERENCES:
1. Stransinger, Susan K., Urinalysis and Body Fluids, Third Edition, F.A. Davis Book
Publisher, 1994, Pages 1 to 10 and 51 to 74.
2. Haber, Meryl H., Urinary Sediment: A Textbook Atlas, American Society of Clinical
Pathologist Book Publisher, 1994.
3. Multistix 10 with SG Package Insert, Bayer Corporation; Diagnostics Division, 1999.
4. Kova Trol: Human Urinalysis Controls Package Insert, Hycor Biomedical Inc., 2001.
5. Manual Urinalysis by Bayer 10-SG Multistix SOP, U.1.1
6. Specific Gravity Determinations by Refractometer SOP, U.5.1
7. Clinitest Determination of Reducing Substances in Urine SOP, U.6.1
8. Acetest Determination of Ketones in Urine SOP, U.8.1
9. Ictotest Determination of Bilirubin in Urine SOP, U.9.1
10. SSA Determination of Protein in Urine SOP, U.10.1
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Appendix 1
SOP VALIDATION
SOP NAME:
U.2. Manual Urinalysis Microscopic Exam
Clear and specific title and principle:
Comments:
yes / no
yes / no
yes / no
yes / no
yes / no
Date: __________________
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Change
QA
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OIC
Med. Dir.
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U.2. Manual Urinalysis Microscopic Exam
Appendix 2
SOP APPROVAL
SIGNATURE
DATE
PREPARER
QA COORDINATOR
LABORATORY OIC
MEDICAL DIRECTOR
ANNUAL REVIEW
REVIEWER SIGNATURE
DATE
REVIEWER SIGNATURE
DATE: ___________
DATE
# COPIES __________
SUPERSEDES:
DATE SOP RETIRED: __________
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Expiration:
Month/Year:
Dipstick Lot#
Expiration:
Date in use:
Supervisor Review:
Dipstick Lot#
Expiration:
Date in use:
Review Date:
Date
Color/
Clarity
Glucose
Bilirubin
Ketones
Specific
Gravity
Blood
pH
Protein
Acceptable
Range:
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
Comments:
Refer to SOPs U.1.1 and U.2.1 for Urine Dipstick and Microscopic QC resulting procedures
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Urobilin.
Nitrate
Leu.
Esterase
WBC/hpf
RBC/hpf
Crystals
Casts
Tech
Initials
Expiration:
Dipstick Lot#
Dipstick Lot#
Date
Color/
Clarity
Glucose
Month/Year:
Expiration:
Date in use:
Supervisor Review:
Expiration:
Date in use:
Review Date:
Bilirubin
Specific
Gravity
Ketones
Blood
pH
Protein
Acceptable
Range:
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
Comments:
Refer to SOPs U.1.1 and U.2.1 for Urine Dipstick and Microscopic QC resulting procedures
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Urobilin.
Nitrate
Leu.
Esterase
WBC/hpf
RBC/hpf
Crystals
Casts
Tech
Initials
_______________________________
Date:
___________________________________
Reagent
Current Lot
Result
Current Lot
Acceptable
Range
Current Lot
Acceptable
Range
Acceptable
Yes /
No
Yes /
No
Casts
Yes /
No
Crystals
Yes /
No
Yes /
No
Yes /
No
Yes /
No
Yes /
No
Yes /
No
Yes /
No
*Bacteria
*Note:
None
None
If bacterial presence is suspected in the quality control material, it is unacceptable for use. Notify the
supervisor immediately.
Yes
No
Comments:
Tech Signature:
Date:
Supervisor review :
Date:
Supervisor Comments:
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Appendix 6 - Patient Manual Urinalysis Result Report
Patient Information:
_____________________________
___________________________________
Patient Name:
ID#
Sample Information:
Date of Birth:
______________________________
Analyte
_____________________
Patient Results
Reference Range
0-1 / LPF
Other Casts:
>
50
0-1
1-3
3-5
5-10
10-25
25-50
0-2
2-5
5-10
10-25
25-50
> 50
0-2 / HPF
0-2
2-5
5-10
10-25
25-50
> 50
0-2 / HPF
0-2
2-5
5-10
10-25
25-50
> 50
0-2 / HPF
None
Few
Trace
Moderat
e
Many
None
Ligh
t
Moderat
e
Few
Moderat
e
Type:
None / LPF
Type:
Bacteria / HPF
Mucus / HPF
None-Few / HPF
Heavy
None-Light / HPF
Crystals / HPF:
None / HPF
Type:
None
Many
None / HPF
Type:
None / HPF
Spermatozoa / HPF:
Yeast / HPF:
Trichomonas
None
Few
Moderat
e
Many
None
Few
Moderat
e
Many
None / HPF
None
Few
Moderat
e
Many
None / HPF
Comments:
Tech Signature:
Supervisor review required for
all critical values.
Required?
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Report Date/Time:
Yes / No
Signature:
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Comments:
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