CAT. NO. 1TP01002-TC40, ITPO1002-DS50 A ADVANCED QUALITY ™ (01.05.03.122 -170702 ONE STEP HBsAb TEST (Serum/ Plasma) FOR IN VITRO DIAGNOSTIC USE ONLY This package insert must be read carefully prior to use. Package insert instructions must be carefully followed. Reliability of assay results cannot be guaranteed if there are any deviations from the instructions in this package insert. 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 or plasma. This test is a rapid test as an aid in the diagnosis of relative infection and relative symptom; it only provides preliminary analysis results but not critical diagnosis criteria, the obtained results should be analyzed in connection with other clinical information, e.g. clinical symptoms, and alternate test to make final decision. 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 TEST uses the “sandwich principle”,' 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 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/mI. REAGENTS AND MATERIALS SUPPLIED FOR CARD TEST, ITP01002-TC40 * 40x Test card individually foil pouched with a desiccant 40 x Plastic dropper 1x Package insert FOR STRIP TEST, ITP01002-DS50 ; | 50 x Test strip individually foil pouched with a desiccant 1 x Package insertMATERIALS REQUIRED BUT NOT PROVIDED * Timer or stopwatch, Precision pipette or similar equipment to deliver 100,L/80pL. Blood collection devices Biohazard disposal container Disposable gloves STORAGE AND STABILITY The kit has a 24 month shelf-life from the date of manufacture. Store the unused kits at 2°C-30°C. If stored refrigerated, ensure that the sealed pouch is brought to room temperature (10°C-30°C) before opening for testing, WARNINGS AND 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 Biomedical Laboratories’ or other equivalent guidelines. 1. For in vitro diagnostic use only. 2. All positive results must be confirmed by an alternate method. 3. Treat all specimens as though potentially infectious. Wear gloves and protective clothing when handling specimens. 4. Clean and disinfect all spills of specimens and reagents using a suitable disinfectant,> such as 1% Sodium Hypochlorite for nonradioactive material” or 2% Glutaraldehyde for spills containing radioactive material.” 5. Devices used for the assay should be sierilized before being disposed. 6. Do not use kit materials beyond their expiration dates. 7. Do not interchange reagents from one kit lot to another. 8. Donot re-use the test cards or any single use accessories. 9. Do not use the test if the foil pouch is damaged. SAMPLE COLLECTION AND STORAGE SERUM OR PLASMA 1. SERUM Use the standard venous phlebotomy procedure to collect a whole blood sample by a tube NOT containing any of the following anticoagulants: EDTA, heparin, or sodium citrate. Leave to settle for 30 minutes for blood coagulation and then centrifuge blood to get serum specimen of supernatant. 2. PLASMA Use the standard venous phlebotomy procedure to collect a whole blood sample by a tube containing any of the following anticoagulants: EDTA, heparin, or sodium citrate. And then centrifuge blood to get a plasma specimen. Note: 1. If serum or plasma specimens are not tested immediately, they should be refrigerated at 2C-8C., For storage period longer than 7days, freezing is recommended. They should be brought to room temperature before testing. 2. Serum or plasma specimens containing a precipitate may yield inconsistent testresult. Such specimens must be clarified before testing. 3, Avoid frequent (more than 3 times) thaw-and-freeze of specimens. 4. Shipped specimens should be packed in compliance with federal and international regulations covering the transportation of etiologic agents. 5. Up to 0.1% of sodium azide can be cimen as preservative without affecting the results of the assay. added to specimen as P' ASSAY PROCEDURE Do not open the pouch until you are ready to perform a test, and the single-use test is suggested to be used under low environment humidity (RHS70%) within 1 hour. FOR TEST CARDS: 1. Bring all reagents and specimens to room temperature. 2. Remove the test from the foil pouch and place on a clean dry surface. 3. Identify the test for each specimen or control. 4. Dispense 100pL (or 3 drops using the plastic dropper provi specimen or control into the sample well on the card. 5. Wait for at least 15 minutes (and 20 minutes at most) to read the result. AS}] [JAS ided in the kit) of the #GcGs Oh Or()s Positve Negatve “Invalid ae — ao SS (7~\ an © ett WS t t FOR TEST STRIPS: to room temperature. 1. Bring all reagents and specimens Remove the test from the foil pouch and place on a clean dry surface. 2. ° f t 3. Identify the test for each specimen or control ; 4 poaly 204 of specimen or control to the sample pad behind the ( {41 ) mark at the bottom of test strip. , 5. Wait for at least TP nutes (and 20 minutes at most) to read the result. etBAB Ae ee ee 22 4 22 wel ele) Le c : A ic} jc tTheWT T TRaT Wi) HE) 80pl of specimen —pe Postive Nogatvo Invalid Caution: Use a clean dropper or pipette for every specimen to avoid cross- contamination. NOTE: 1. A positive result may be interpreted early, however read any negative at 15 minutes to ensure sample is negative and not a low concentration of HBsAb. Do not interpret the result after 20 minutes. 2. The positive results could appear as soon as 1 minute for a sample with high levels of HBsAb. 3. _ No test provides absolute assurance that a specimen does not contain low levels of HBsAb such as those present at a very early stage of infection. A negative result does not preclude the possibility of exposure to or infection with HBV virus. INTERPRETATION OF THE TEST 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. 1. Positive: Both purplish red test band and purplish red control band appear on the membrane. 2. Negative: Only the purplish red control band appears on the membrane. The absence of a test band indicates a negative result. 3. Invalid: There should always be a purplish red control band in the control region regardless of test result. If control band is not seen, the test is considered invalid, Repeat the test using a new test device. PERFORMANCE CHARACTERISTICS The Advanced Quality™ ONE STEP HBsAb TEST showed equivalent detectability forHBsAb with EIA immunoassay. HBsAb Concentration as low as 10mlU/mI i in human serum is detectable. A result of 99% correlation to EIA t i um study of 1300 specimens, lest was determined by a clinical LIMITATIONS 1. en BIBI 1. False reactive results may be obtained with i , , of two types. with any diagnostic test and usually consist NONSPECIFIC REACTIVES immunoassay system has the potential for nonspecific reaction in human serum or plasma. 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. The assay should be performed in normal room temperature. Test cards/strips should be used immediately after being taken from the package. Avoid exposing the test cards/strips in the air for too long before use. Only samples that are not hemolyzed and that are with good fluidity can be used in this test. Fresh samples are best but refrigerated and frozen samples can also be used after thawing and balancing to the room temperature(10’C-30°C). Do not agitate the sample. Insert a pipette just below the surface of the sample to collect the specimen. lY Poeana 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, | | 5 ; a U. S. Department of Health and Human Services. Biosafety in microbilogical and biomediesl laboratories. HHS Publication (NIH) 88-8395. Washington: U.S. inti May 1988. raed pl eae Laboratory biosafety manuel. Geneva. World Health Organization, 1983. 4 liar jinical Laboratory Standards. Protection of laboratory ma cor ecious dieeas0 transmitted by blood, body fluids, and tissue: eet trae ec NGeLS Document M29-T. Villanova, PA.: NCCLS, 1989 es Disease Control. Recommendation for prevention of HIV Cawley Centers for me setting. MMWR 36, Supplement No. 2S, 1987. a sree ea inger F. B., Dreesman. G. R., and Melnick, J. L., Immunological ehulster, L. M., Hol VF. Ba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-higl h 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. Millman, |, and McMichael, JC, Glycoprotein of natural origin with an affinity for hepatitis B surface antigen. Infection and Immunity. 21:879-885. 1978. nad InTec PRODUCTS, INC. 332 Xinguang Road, Xinyang Ind. Area, Haicang, Xiamen, 361022, P.R. China Tel: +86 592 6807188 Email: intecproducts@asintec.com www.intecasi.com