S02 ATTACHMENT D.16 - FAV_SEROLOGY_11_QUALITATIVE_PREGNANCY_TEST.pdf

FAV Serology 11 Sure-Vue HCG-STAT Qualitative Pregnancy Test

Copy of version 1.0 (approved and current)

Last Approval or Periodic Review Completed

6/17/2022

Next Periodic Review Needed On or Before

6/17/2024

Effective Date 2/12/2018

Controlled Copy ID 26036

Location No Physical Location

Organization Veterans Health System of the Ozarks

Author Shelly Stewart

Approval and Periodic Review Signatures

Type Description Date Version Performed By Notes

Approval Lab Director 6/17/2022 1.0 Constantine Theoharis

Periodic review

Designated Review er

3/9/2022 1.0 Patricia Gregg

Periodic review

Designated Review er

3/7/2022 1.0 Debbie Gilbert-Hardy Recorded w hen document added to MediaLab

Approval Lab Director 2/28/2020 1.0 Dr. K Alexander Recorded w hen document added to MediaLab

Approvals and periodic reviews that occured before this document was added to the MediaLab Document Control system may not be listed.

Version History

Version Status Type Date Added Date Effective Date Retired

1.0 Approved and Current First version in Document Control 3/7/2022 2/12/2018 Indefinite

SURE-VUE hCG-STAT

QUALITATIVE PREGNANCY TEST

Principle

Sure-Vue Serum/Urine hCG-STAT is a rapid chromatographic immunoassay for the qualitative detection of human chorionic gonadotropin in serum or urine to aid in the early detection of pregnancy.

Human chorionic gonadotropin (hCG) is a glycoprotein hormone secreted by the developing placenta shortly after conception. In normal pregnancy, hCG can be detected in urine or serum as early as 7-10 days following conception. hCG levels continue to rise very rapidly, frequently exceeding 100 mIU/ml by the first missed menstrual period and peaking in the 100,000 to 200,000 mIU/ml range about 10-12 weeks into pregnancy. The appearance of hCG soon after conception and its subsequent rise in concentration during early gestational growth make it an excellent marker for the early detection of pregnancy.

Sure-Vue Serum/Urine hCG-STAT is a rapid test that qualitatively detects the presence of hCG at the sensitivity of 10 mIU/mL in serum and 20 mIU/mL is urine. The test utilizes a combination of monoclonal and polyclonal antibodies to selectively detect elevated levels of hCG in serum or urine. At the level of claimed sensitivity, the Sure-Vue Serum/Urine hCG-Stat shows no cross-reactivity interference from the structurally related glycoprotein hormones hFSH, hLH and hTSH at high physiological levels.

The test utilizes a combination of antibodies including mouse monoclonal anti-hCG antibodies and goat polycolonal anti-hCG antibodies to selectively detect elevated levels of hCG. The assay is conducted by adding serum or urine specimen to the specimen well of the test device and observing the formation of colored lines. The specimen migrates via capillary action along the membrane to react with the colored conjugate. Positive specimens react with the specific colored antibody conjugates and form a colored line at the test line region of the membrane. Absence of this colored line suggests a negative result. To serve as a procedural control, a colored line will always appear at the control line region if the test has been performed properly.

Specimens

A. Specimen Preparation There is no special specimen preparation.

B. Specimen type and collection

1. Blood should be collected (see Phlebotomy Manual) and the serum separated by standard procedures. Either a red/black (no gel, clot

Approved and current. Effective starting 2/12/2018. FAV Serology 11 (version 1.0) Sure-Vue HCG-STAT Qualitative Pregnancy Test

Controlled copy ID 26036. Printed on 3/14/2023 10:20 AM (EDT). Page 1 of 12 activator) or red/yellow (gel) tube may be used. Centrifuge the serum as soon as guidelines suggest to avoid hemolysis. Use clear non-hemolyzed specimens when possible.

2. A urine specimen must be collected in a clean and dry container. A first morning specimen is preferred since it generally contains the highest concentration of hCG; however, urine specimens obtained at any time of the day may be used. Urine specimens exhibiting visible precipitation should be centrifuged and the clear supernatant used for testing.

C. Specimen rejection criteria

1. Do not use grossly hemolyzed specimens.

2. Specimens must contain the correct labeling (see the Phlebotomy

Manual for Patient Identification procedure). All mislabeled specimens must be entered into the Specimen Rejection and Incident Log and recollected.

3. See Appendix - Specimen Rejection Criteria found in the Specimen Collection manual for the other causes of rejection.

D. Specimen Stability

1. If specimens cannot be tested on the day of collection, store them at 2-8º C for up to 48 hours.

2. If longer that 2 day storage is required, freeze specimens at -20° C or below for 2 months. Mix specimens after thawing prior to testing.

Avoid successive freezing and thawing.

E. Specimen Transportation

1. Blood specimen should be transported to lab at room temperature

(20º - 25º C). See the Phlebotomy Manual and the Infection Control Policy in the General Policy and Procedure Manual for correct transportation procedure when transporting specimens to the laboratory.

2. Off-site specimens should be centrifuged and serum separated.

Transport at 2 – 8 º C (refrigerated). See the Phlebotomy Manual and the Infection Control Policy in the Laboratory General Policy and Procedure Manual for correct transportation procedure when transporting specimens to the laboratory.

Materials and Reagents

A. Reagents

1. Test device- contains anti-alpha hCG gold conjugate and anti-hCG coated on the membrane.

2. Sure-Vue hCG STAT Serum Control Set:

Approved and current. Effective starting 2/12/2018. FAV Serology 11 (version 1.0) Sure-Vue HCG-STAT Qualitative Pregnancy Test

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a. 250 mIU/mL hCG Positive Serum Control - contains normal human serum and sodium azide 0.2%

b. 10 mIU/mL Positive Serum Control contains normal human serum and sodium azide 0.2%

c. Negative Serum Control - contains sodium azide 0.2%. Serum control materials are manufactured from source plasma. Each donor unit plasma used in the preparation was tested and found negative for HbsAG, HIV 1 and 2, HIV-lag and HCV by FDA approved methods

3. Sure-Vue hCG Urine Control Set:

a. 250 mIU/ml hCG Positive Urine Control – contains normal human urine and sodium azide 0.2%

b. 25 mIU/ml hCG Positive Urine Control – contains normal human urine and sodium azide 0.2%

c. Negative Urine Control – contains sodium azide 0.2%

B. Materials

1. Disposable dropper pipettes

2. Timer

Storage and Stability

Store test kit as packaged in the sealed pouch at room temperature (2° - 30ºC). Do not freeze. Do not use the test kit beyond expiration date.

Unopened controls are stable until the expiration date on the label when stored refrigerated at 2° - 8ºC. Do not freeze. Do not use the controls if they are cloudy or if precipitates are observed in the vials.

This may be an indication of reagent instability or deterioration. Do not use controls beyond the expiration date.

The failure of the positive or negative control to produce a positive or negative result respectively may indicate reagent instability or deterioration of the controls.

Quality Control

Internal Procedural Controls.

Internal procedural controls are included in the test. A red line appearing in the control region (C) is the internal procedural control. It confirms sufficient specimen volume and correct procedural technique. A clear background is an internal negative background control. If the test is working properly, the background in the result area should white to light pink and not interfere with the ability to read the test result.

Record the Internal Controls on the Workload list and in the Reagent Logbook.

Approved and current. Effective starting 2/12/2018. FAV Serology 11 (version 1.0) Sure-Vue HCG-STAT Qualitative Pregnancy Test

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External Quality Control

Perform external quality control consisting of a negative, low positive and positive control under the following circumstances:

1. With every new lot of reagent kits (urine and serum controls)

2. With every new shipment of reagent kits, regardless of the lot number (a new shipment of the same lot number still requires external controls) (urine and serum controls)

3. Every 30 days (urine and serum controls)

4. Troubleshooting

5. With each new operator.

Serum and Urine controls are run with a new lot and new shipment of reagent kits plus every 30 days.

Allow the controls to reach room temperature prior to testing. The controls are ready to use. No dilution is required. Thoroughly mix the contents of each bottle by gentle swirling. All QC results are entered into the LIS (Manually Order and Accession QC). The positive controls should yield positive results and are entered as “1”. The negative control should yield negative results and is entered as “0”. The failure of the positive or negative control to produce a positive or negative result, respectively, may indicate reagent instability or deterioration of the controls. Record all corrective action in the Reagent Logbook. See Individualized Quality Control Plan for any QC failures. The IQCP may need to be reassessed and a new IQCP signed.

Troubleshooting steps include:

Check procedure for proper performance of test Repeat testing Open a new box of testing devices and repeat testing and QC Contact Sure-Vue technical support Report the problem to the lead technologist or supervisor

No patient results should be reported with invalid control results.

Procedure

1. Both test device and patient samples and controls should be brought to room temperature 20-30º C prior to testing. Testing devices are kept normally at room temperature and are ready to use. Controls are refrigerated and must be brought to room temperature prior to use.

2. Remove the test device from its protective pouch and use it as soon as possible. Label the device with patient or control identification.

3. Place the test device on a clean and level surface. Holding the dropper in a vertical position, dispense 3 full drops (approximately 100 ul) of the specimen or control into the sample well of the test device. For each

Approved and current. Effective starting 2/12/2018. FAV Serology 11 (version 1.0) Sure-Vue HCG-STAT Qualitative Pregnancy Test

Controlled copy ID 26036. Printed on 3/14/2023 10:20 AM (EDT). Page 4 of 12 sample or control, use a separate pipette and device. Start the timer: 5 minutes for serum or 3 minutes for urine.

4. Wait for colored bands to appear. It is important that the background is clear before the result is read. Depending on the concentration of hCG, positive results may be observed in as short as 40 seconds. However, to confirm negative results, the complete reaction time (3 or 5 minutes) is required. If a control line fails to appear, the testing is invalid. If the background fails to clear making the reading of the test difficult, the testing is invalid.

Note: A low hCG concentration might result in a weak line appearing in the test region (T) after an extended period of time. See below for interpretation.

Interpretation of Results

Negative: One red line appears in the control region (C). No apparent red or pink line appears in the test region (T).

Positive: Two distinct red lines appear. One line should be in the control region

(C) and another line should be in the test region (T).

Uninterpretive: A very faint pink line in the test region (T) and a red line in the control region(C) should be reported as “indeterminant”. The serum specimen should be referred for Beta-HCG testing.

Note: A sample hCG concentration below the low cut-off level of this test might result in a weak line appearing in the test region (T) after an extended period of time. Because of the high sensitivity of this test, urine results should be read between 3 and 5 minutes only. Serum results should be interpreted between 5 and 7 minutes only. A result seen after these times could be indicative of a low hCG level in the sample. If such results are seen, it is recommended that the test be repeated with a new sample in 48 – 72 hours or that an alternate confirmation method is used.

Note: The intensity of the red color in the test line region (T) will vary depending the concentration of hCG present in the specimen. However, neither the quantitative value nor the rate of increase in hCG can be determined by this qualitative test.

Invalid: Control line fails to appear. Insufficient specimen volume or incorrect procedural techniques are the most likely reasons for control line failure. Review the procedure and repeat the test with a new test device. External quality control, with urine or serum controls, may be run to determine if deterioration of the kit has occurred. A new lot of testing devices may also be opened for repeat testing. If the problem persists, discontinue using the test kit. Contact the lead technologist or supervisor.

Approved and current. Effective starting 2/12/2018. FAV Serology 11 (version 1.0) Sure-Vue HCG-STAT Qualitative Pregnancy Test

Controlled copy ID 26036. Printed on 3/14/2023 10:20 AM (EDT). Page 5 of 12

All results are recorded on the workload list (including internal controls) and entered into the LIS. Also, place in the Reagent Logbook for monthly review by supervisor/lead technologist. Place the workload list in serology for review.

Expected Values

Negative results are expected in healthy non-pregnant women and healthy men.

Healthy pregnant women have hCG present in their urine and serum specimens.

The amount of hCG will vary greatly with gestational age and between individuals.

The Sure-Vue Serum/Urine hCG STAT has a sensitivity of 10 mIU/mL in serum and 20 mIU/mL in urine and can detect pregnancy as early as 1 day after the first missed menses.

Performance Characteristics

Accuracy

A multi-center clinical evaluation was conducted comparing the results obtained using Sure-Vue ® Serum/Urine hCG STAT and another commercially available serum/urine membrane hCG test. The urine study included 100 specimens and both assays identified 50 negative and 50 positive results. The serum study included 100 specimens and both assays identified 50 negative and 50 positive results. The results demonstrated 100% overall agreement (for an accuracy of >99%) of the Sure-Vue Serum/Urine hCG-STAT when compared to the other Serum/Urine membrane hCG test.

Sensitivity and Specificity

Sure-Vue Serum/Urine hCG-STAT detects hCG at concentrations of 10 mIU/mL or greater in serum and 20 mIU/mL or greater in urine. The test has been standardized to the W.H.O. Fourth International Standard (75/589). The addition of LH (300 mIU/mL), FSH (1,000 mIU/mL), and TSH (1,000 µIU/mL) to negative (0 mIU/mL hCG) and positive (10 mIU/mL hCG in serum/20 mIU/mL hCG in urine) specimens showed no cross-reactivity.

Approved and current. Effective starting 2/12/2018. FAV Serology 11 (version 1.0) Sure-Vue HCG-STAT Qualitative Pregnancy Test

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Interfering Substances

The following potentially interfering substances were added to hCG negative and positive specimens.

All substances listed in mg/mL unless otherwise noted.

Acetaminophen 20 Cocaine 10 Ibuprofen 20 Acetone 1,000 Codeine 10 Methadone 10 Acetylsalicylic Acid 20 Cholesterol 500 Methamphetamine 10 Acetoacetic Acid 2,000 Creatine 20 Methanol 10% Ampicillin 20 Dextromethorphan 20 Morphine 0.6 Ascorbic Acid 20 DMSO 5% Oxalic Acid 40 Atropine 20 EDTA 80 Phenothiazine 20 Albumin 2,000 Ephedrine 20 Phenylpropanolamine 20 Β-Hydroxybutyrate 2,000 Ethanol 1% Pregnanediol 2 salt Benzoylecgonine 10 Estriol 2 Salicylic Acid 20 Bilirubin 20 Estrone 3-Sulfate 10 Tetracycline 20 Brompheniramine 20 Gentisic Acid 20 Triglyerides 1,200 Caffeine 20 Glucose 2,000 Theophylline 20 Cannabinol 10 Hemoglobin 1,000 Urea 2,000 Clomiphene 100 Heroin 1 Uric Acid 20

None of the substances at the concentrations tested interfered in the assay.

Downtime

If in a rare circumstance the lab is unable to perform a rapid qualitative pregnancy test, a serum Beta-HCG can be performed.

Limitations

1. Very dilute urine specimens, as indicated by a low specific gravity, may not contain representative levels of hCG. If pregnancy is still suspected, a first morning urine specimen should be collected 48 hours later and tested.

2. False negative results of hCG are below the sensitivity level of the test.

When pregnancy is still suspected, a first morning urine or serum specimen should be collected 48 hours later and tested.

3. Very low levels of hCG (less than 50 mIU/mL) are present in urine and serum specimen shortly after implantation. However, because a significant number of first trimester pregnancies terminate for natural reasons, a test result that is weakly positive should be confirmed by retesting with first morning urine or serum specimen collected 48 hours later.

Approved and current. Effective starting 2/12/2018. FAV Serology 11 (version 1.0) Sure-Vue HCG-STAT Qualitative Pregnancy Test

Controlled copy ID 26036. Printed on 3/14/2023 10:20 AM (EDT). Page 7 of 12

4. This test reliably detects intact hCG up to 500, 000 mIU/mL. It does not reliably detect hCG degradation products, including free-beta hCG and beta core fragments. Quantitative assays used to detect hCG may detect hCG degradation products and therefore may disagree with the results or this rapid test.

5. A number of conditions other than pregnancy, including trophoblastic disease and certain non-trophoblastic neoplasms including testicular tumors, prostate cancer, breast cancer, and lung cancer, can cause elevated levels of hCG. Therefore, the presence of hCG in serum or urine should not be used to diagnose pregnancy unless these conditions have been ruled out.

6. As with any assay employing mouse antibodies, the possibility exists for interference by human anti-mouse antibodies (HAMA) in the specimen.

Serum samples from patients who have undergone therapy or diagnostic procedures that use preparations of Mouse Monoclonal Antibodies may contain Human Anti-Mouse Antibodies (HAMA). The antibodies in these specimens may produce false results, both positive and negative.

7. This test provides a presumptive diagnosis for pregnancy. A confirmed pregnancy diagnosis should only be made by a medical provider after all clinical and laboratory findings have been evaluated.

References

Sure-Vue Serum/Urine hCG-STAT package insert. DN: 1155810104, Effective Date 2014-05-27

Approved and current. Effective starting 2/12/2018. FAV Serology 11 (version 1.0) Sure-Vue HCG-STAT Qualitative Pregnancy Test

Controlled copy ID 26036. Printed on 3/14/2023 10:20 AM (EDT). Page 8 of 12

Sure-Vue Serum/Urine HCG Kit

Parallel Testing Form

Parallel Testing

New Kit Lot #

Old Kit Lot #

Serum High Pos Control

Lot #

Serum Low Pos Control

Lot #

Serum Negative Control

Lot #

Urine High Pos Control

Lot #

Urine Low Pos Control

Lot #

Urine Negative Control

Lot #

Expires Expires Expires Expires Expires Expires Expires Expires

New Kit Results

Serum Hi Pos Control

Serum Lo Pos Control

Serum Neg Control

Urine Hi Pos Control

Urine Lo Pos Control

Urine Neg Control

Old Kit Results

Serum Hi Pos Control

Serum Lo Pos Control

Serum Neg Control

Urine Hi Pos Control

Urine Lo Pos Control

Urine Neg Control

Corrective Action:

Parallel Testing Acceptable: YES NO

Date/Tech: Reviewed:

Approved and current. Effective starting 2/12/2018. FAV Serology 11 (version 1.0) Sure-Vue HCG-STAT Qualitative Pregnancy Test

Controlled copy ID 26036. Printed on 3/14/2023 10:20 AM (EDT). Page 9 of 12

Sure-Vue Serum/Urine HCG Kit 30 Day QC Form

30 Day QC – Serum and Urine

HCG Kit Lot # Expires

Serum High Pos Control Lot # Expires

Serum Low Pos Control Lot # Expires

Serum Negative Control Lot # Expires

Urine High Pos Control Lot # Expires

Urine Low Pos Control Lot # Expires

Urine Negative Control Lot # Expires

Serum Hi Pos QC results

Serum Lo Pos QC results

Serum Negative QC results

Urine Hi Pos QC results

Urine Lo Pos QC results

Urine Negative QC results

Corrective Action:

30 Day QC Acceptable: YES NO

Date/Tech: Reviewed:

Approved and current. Effective starting 2/12/2018. FAV Serology 11 (version 1.0) Sure-Vue HCG-STAT Qualitative Pregnancy Test

Controlled copy ID 26036. Printed on 3/14/2023 10:20 AM (EDT). Page 10 of 12

HCG SERUM AND URINE CONTROLS PARALLEL TESTING FORM

SERUM URINE

NEW LOT HCG HIGH POSITIVE CONTROL LOT NUMBER:

EXPIRATION DATE:

NEW LOT HCG LOW POSITIVE CONTROL LOT NUMBER:

EXPIRATION DATE:

NEW LOT HCG NEGATIVE CONTROL LOT NUMBER:

EXPIRATION DATE:

HIGH POSITIVE

CONTROL

LOW POSITIVE

CONTROL

NEGATIVE CONTROL

OLD LOT HCG HIGH POSITIVE CONTROL LOT NUMBER:

EXPIRATION DATE:

OLD LOT HCG LOW POSITIVE CONTROL LOT NUMBER:

EXPIRATION DATE:

OLD LOT HCG NEGATIVE CONTROL LOT NUMBER:

EXPIRATION DATE:

HIGH POSITIVE

CONTROL

LOW POSITIVE

CONTROL

NEGATIVE CONTROL

HCG KIT LOT NUMBER:

EXPIRATION DATE:

CORRECTIVE ACTION:

PARALLEL TESTING ACCEPTABLE: YES NO

DATE/TECH INITIALS: REVIEWED:

Approved and current. Effective starting 2/12/2018. FAV Serology 11 (version 1.0) Sure-Vue HCG-STAT Qualitative Pregnancy Test

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Table 1 Test Caption

Table 2 Biennial Review

Table 3 Changes

Implementation Date:

Revision Supersedes: Sure-Vue hCG-STAT Version SR 2017-03

Location of Original Signed Copy: Microbiology Procedure Manual

Copies Located: VHSO Intranet (Sharepoint Sites-P&LMS) – Serology Procedure Manual

Written by (Signature): Date:

Shelly M. Stewart, MT(ASCP)SLSCM

Reviewed by (Signature): Date:

Patrick Quinlan, MLS(ASCP)CM

Approved by (Signature): Date:

Stephen A. Schichman, MD, PhD, Laboratory Director, P&LMS

Associated Documents:

Date Review by (Signature) Findings of Review No changes Minor changes-see below Revision initiated

No changes Minor changes-see below Revision initiated

No changes Minor changes-see below Revision initiated

No changes Minor changes-see below Revision initiated

No changes Minor changes-see below Revision initiated

No changes Minor changes-see below Revision initiated

Date Page # Description of Change 2/10/18 1,2 Corrected specimen type to blood

2/10/18 4 Serum QC added to 30 day QC testing; edited QC forms to reflect this change

5/7/21 9,10 Reformatted QC form attachments

Controlled Version GN 2018-01

Approved and current. Effective starting 2/12/2018. FAV Serology 11 (version 1.0) Sure-Vue HCG-STAT Qualitative Pregnancy Test

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