Declaration: As with all diagnostic tests, a definitive clinical diagnosis should not be based on the result of a single test, but should only be made by the physician after all clinical and laboratory findings have been evaluated.
SPECIMEN COLLECTION AND PREPERATION
( SERUM / WHOLE BLOOD )
Catalog No. Product Name
ABT-IDT-A7 HBsAb Serum Strip
ABT-IDT-B7 HBsAb Serum Cassette
ABT-IDT-A79 HBsAb Whole Blood Strip
ABT-IDT-B79 HBsAb Whole Blood Cassette
Accu-Tell ® Rapid HBsAb Serum/Whole Blood Test is a rapid, immunochromatographic assay for the detection of antibodies to Hepatitis B Surface Antigen (HBSAB) in human whole blood, serum or plasma. The test provides a visual, qualitative result. It is for professional use.
Accu-Tell ® Rapid HBsAb Serum/Whole Blood Test uses the “sandwich principle”,1 a solid phase colloidal gold enhanced immunoassay technique for determination of HBsAb in human whole blood, serum or plasma.
SERUM: 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.
WHOLE BLOOD: The nitrocellulose membrane is coated with HBsAg in the test region and anti-HBsAg antibody in the control 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. An HBsAb positive specimen produces a distinct color band in the test region, formed by the specific antibody-HBsAg complex. Absence of this colored band in the test region suggests a negative result. A colored band always appears in the control region serving as procedural control regardless of the test result. The sensitivity of the test is 10mIU/ml.
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 Biomedical Laboratories1 or other equivalent guidelines.2-4
1. For in vitro diagnostic use only.
2. All serum or plasma specimens should be treated as infectious material. Do not contact the test device without wearing safety gloves.
3. Clean and disinfect all spills of specimens using a suitable disinfectant5 such as 1% Sodium Hypochlorite for nonradioactive material6 or 2% Glutaraldehyde for spills containing radioactive material.7
4. Devices used for the assay should be sterilized before being disposed.
5. Do not use the test device or reagent beyond expiration date.
SPECIMEN COLLECTION AND PREPERATION
1. Either serum or plasma may be used in the test.
2. 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.
3. 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.
4. If specimen are to be stored, they should be refrigerated at 2-8oC . For long term storage(over 3 days), the specimens should be frozen.
5. If specimens are to be shipped, they should be packed in compliance with federal regulation covering the transportation of etiologic agents.
6. Avoid frequent (more than 3 times) thaw-and-freeze of specimens.
7. Up to 1% of sodium azide can be added to specimen as preservative without affecting the results of assay.
SPECIMEN COLLECTION AND PREPERATION
Whole Blood:
1. Collect whole blood specimens following regular clinical laboratory procedures.
2. Heparinized capillary tubes must be used for collecting whole blood samples. Do not use hemolyzed blood samples.
3. Whole blood specimens should be used immediately after collection.
Serum or Plasma
1. Collect serum or plasma specimens following regular clinical laboratory procedures.
2. Only those specimens that are clean, clear and with good fluidity can be used for the assay.
3. Those specimens that are apparently hemolyzed, extremely thickened or with very high fat level are NOT suitable for the assay.
4. Storage: A specimen should be refrigerated if not used the same day of collection. Specimens should be frozen if not used within 3 days of collecting. Avoid freezing and thawing the specimens more than 2-3 times before use. 0.1% of sodium azide can be added to specimen as preservative without affecting the results of the assay.
For Test Cassette:
1. Remove the test cassette from the foil pouch and place on a clean dry surface.
2. Identify the test cassette for each specimen or control.
3. Dispense 100μl (3drops) of the specimen or control into the sample well on the card.
4. Interpret test results at 15 minutes.
For Test Strip:
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 80μl of specimen to the sample pad behind the ( ↓↓↓ ) mark at the bottom of test strip.
5. Interpret test results at 15 minutes.
Caution: Use a disposable pipette tip for each transfer to avoid cross-contamination
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.
Positive: In addition to the control band, a purplish red test band also appears on the test region.
Negative: Only one purplish red test band appears on the control region.
Invalid: Neither test band nor control band appears. The specimen should be tested again.
False reactive results may be obtained with any diagnostic test and usually consist of two types.
1. NONSPECIFIC REACTIVES
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 this hepatitis B binding substance is not antibody.9 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.
Accu-Tell ® Rapid HBsAb Test can detect antibodies against HBsAg (HBsAb) in human whole blood, serum, or plasma at concentration as low as 10mIU/ml. The HBsAb detectability is equivalent to that of EIA test. A result of 99% concordance to EIA test was determined by a clinical study of 1300 specimens.
1. Peters, RL. Collins, MJ, O’Beirne, 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.
The above information is just for reference. The actual technical specifications are subject to the insert provided with the product.
Recent Comments