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REV. 51215





FOR IN VITRO DIAGNOSTIC USE ONLY INTENDED USE THE ADVANCED QUALITY ONE STEP HBSAB TEST IS A RAPID, ONE STEP, IMMUNOCHROMATOGRAPHIC ASSAY FOR THE DETECTION OF ANTIBODIES TO HEPATITIS B SURFACE ANTIGEN (HBSAB) IN HUMAN SERUM. THE TEST PROVIDES A VISUAL, QUALITATIVE RESULT, AND IS INTENDED FOR PROFESSIONAL USE. SUMMARY AND PRINCIPLE OF THE ASSAY The Advanced Quality One Step HBsAb Antibody (HBsAb) Test uses the sandwich principle, 1 a solid phase colloidal gold enhanced immunoassay technique for determination of HBsAb in human serum or plasma. The nitrocellulose membrane was immobilized with HBsAg on the test band region and anti-HBsAg antibody on the control band region. During the assay, the specimen is allowed to react with the colored conjugate (HBsAg colloid gold conjugate); the mixture then migrates chromatographically on the membrane by the capillary action. For a positive result, a color band with the specific antibody-HBsAg complex will form on the membrane. Absence of this colored band in the test band region suggests a negative result. To serve as a procedural control, a colored band at control region always appears in the test area. The reagent contains scavenge antibodies to reduce nonspecific reactivity in human serum or plasma specimens. The sensitivity of the test is 10mIU. STORAGE CONDITION The test kits must be stored at 2-30in the sealed pouch and under dry conditions. PRECAUTIONS It is recommended that all specimens be handled in accordance with Biosafety Level 2 practices as described in the CDC NIH Publication, Biosafety in Microbiological and 2 3-4 Biomedical Laboratories or other equivalent guidelines. 1. For in vitro diagnostic use only. 2. All serum or plasma specimens should be treated as infectious material. Do not contact the test card without wearing safety gloves. 3. Clean and disinfect all spills of specimens and reagents using a suitable disinfectant,5 such as 1% Sodium Hypochlorite for nonradioactive material6 or 2% 7 Glutaraldehyde for spills containing radioactive material. 4. Devices used for the assay should be sterilized before being disposed. 5. Do not use beyond expiration date. SPECIMEN COLLECTION AND PREPERATION FOR ANALYSIS 1. Either serum or plasma may be used in the test.

4. 5. 6. 7.

Anticoagulants typically used for blood collection do not interfere with this test. Remove the serum or plasma from the clot or red cells respectively, as soon as possible to avoid hemolysis. Specimens that are apparently hemolyzed, extremely thickened or with very high fat level are NOT suitable for the assay. Specimens containing particulate matter may give inconsistent results and should be clarified before testing. o If specimens are to be stored, they should be refrigerated at 2-8 C. For long term storage (over 3 days), the specimens should be frozen. If specimens are to be shipped, they should be packed in compliance with federal regulation covering the transportation of etiologic agents. Avoid frequent (more than 3 times) thaw-and-freeze of specimens. Up to 1% of sodium azide can be added to specimen as preservative without affecting the results of assay.

MATERIAL PROVIDED Test cards/ test strips, individually foil pouched with a desiccant Package Insert Sample dispensing plastic dropper with each test pouch.(for card only) MATERIALS REQUIRED BUT NOT PROVIDED 1. Precision pipette or similar equipment to deliver 100l 2. Distilled water 3. Positive and negative controls ASSAY PROCEDURE Do not open pouch until you are ready to test the sample. For test cards: 1. Bring all reagents and specimens to room temperature. 2. Remove the test card from the foil pouch and place on a clean dry surface. 3. Identify the test card for each specimen or control. 4. Dispense 100l (3drops) of the specimen or control into the sample well on the card. 5. Interpret test results at 15 minutes.




100ul of specimen (3 drops)

For test strips: 1. Bring all reagents and specimens to room temperature. 2. Remove the test strip from the foil pouch and place on a clean dry surface. 3. Identify the test strip for each specimen or control. 4. Apply at least 80l of specimen to the sample pad behind the ( ) mark at the bottom of test strip. 5. Interpret test results at 15 minutes.

Nonspecific reaction may result from cross-reactions in the immune sandwich complex. Nonspecific reactions may include reactions with certain glycoproteins, such as concanavalin A, 8 which interact with HBsAg. Miliman and McMichaeel have shown that 9 this hepatitis B binding substance is not antibody. Any highly sensitive immunoassay system has the potential for nonspecific reaction in human serum or plasma. 2. NONREPEATABLE REACTIVES Nonrepeatable reactive specimens, as the name implies, test nonreactive upon repeat. This phenomenon is highly dependent upon technique. The most common sources of such nonrepeatable reactive is cross-contamination of non-reactive specimens caused by transfer of residual droplets of high titer, antibody-containing sera on the pipetting device. PERFORMANCE CHARACTERISTICS The Advanced Quality One Step HBsAb Test showed equivalent detectability for HBsAb with EIA immunoassay. HBsAb concentration as low as 10mIU/ml in human serum is detectable. A result of 99% correlation to EIA test was determined by a clinical study of 1300 specimens. BIBLIOGRAPHY 1. Peters, RL. Collins, MJ, OBeirne, AJ, Howton, PA, Hourihan, SL, and Thomas, SF, Enzyme-linked immunosorbent assay for detection of antibodies to murine hepatitis virus. J. Clin. Microbiol. 10:595-597, 1979. 2. U. S. Department of Health and Human Services. Biosafety in microbilogical and biomedical laboratories. HHS Publication (NIH) 88-8395. Washington: U.S. Government Printing Office, May 1988. 3. World Health Organization. Laboratory biosafety manual. Geneva. World Health Organization, 1983. 4. National Committee for Clinical Laboratory Standards. Protection of laboratory workers from infectious disease transmitted by blood, body fluids, and tissue: Tentative guideline. NCCLS Document M29-T. Villanova, PA.: NCCLS,1989 5. Cawley Centers for Disease Control. Recommendation for prevention of HIV transmission in health care setting. MMWR 36, Supplement No. 2S, 1987. 6. Sehulster, L. M., Hollinger, F. B., Dreesman. G. R., and Melnick, J. L., Immunological and biophysical alteration of Hepatitis B virus antigens by sodium hypochlorite disinfection. Appl. And Envir. Microbiol., 42:762-767, 1981. 7. Bond, W. W., Favero. M. S., Peterson, N. J., and Ebert, J. W., Inactivation of Hepatitis B virus by intermediate-to-high level disinfectant chemicals. J. Clin. Microbiol., 18:535-538m 1983. 8. Cawley, LP, Reaction between concanavalin A and the Australia antigen. Am. J. Clin. Pathol. 75:.253, 1972. 9. Millman, I, and McMichael, JC, Glycoprotein of natural origin with an affinity for hepatitis B surface antigen. Infection and Immunity. 21:879-885. 1978.




Caution: Use a disposable pipette tip for each transfer to avoid crosscontamination It is recommended to run a known positive control and negative control in each performance to ensure the assay procedure. INTERPRETATION OF RESULTS The presence or absence of HBsAb provides useful information on the status of individuals with type B viral hepatitis. A positive test for anti-HBsAg antibody can be a useful adjunct for assessing immunity of clinical recovery of the patient. The quantitative measurement of HBsAb is valuable in assessing the level of the immune response to the HBV vaccine. Do not interpret the results after 20 minutes. 1. 2. 3. Negative: Only one purplish red test band appears on the control region. Positive: In addition to the control band, a purplish red test band also appears on the test region. Invalid: Neither test band nor control band appears. The specimen should be tested again.

LIMITATIONS False reactive results may be obtained with any diagnostic test and usually consist of two types. 1. NONSPECIFIC REACTIVES