Rapid test for the qualitative detection of hepatitis B surface antigen (HBsAg) in human serum, plasma, or whole blood.

Transcription

1 REF B - en - 26/11 VIKIA HBsAg Rapid test for the qualitative detection of hepatitis B surface antigen (HBsAg) in human serum, plasma, or whole. SUMMARY AND EXPLANATION The VIKIA "HBsAg" test device is a rapid test, based on the immunochromatography technique (ICT or lateral flow) for the qualitative detection of HBs antigen in human serum, plasma, or whole. The hepatitis B virus is responsible for acute and chronic hepatitis infections. Acute hepatitis can be asymptomatic or present symptoms of varying severity which may progress to fulminant hepatitis in.1 to.5% of cases. Chronicicity occurs in 5 to 1% of cases in adults, but up to 9% of cases in infants following perinatal transmission. Currently, approximately 3 million people worldwide are chronic carriers of the virus (1). Chronic hepatitis B may be asymptomatic and lead to liver lesions of varying severity, possibly evolving to cirrhosis, with an evolution in 5% of cases to hepatocellular carcinoma (2). The hepatitis B virus can be transmitted by parenteral or perinatal pathways or through sexual contact. Persons most at risk are health workers, drug addicts, those with multiple sexual partners, multiple transfusion or hemodialysis patients, as well as close friends and family of an infected subject, and newborns of an infected mother (2). The discovery of the Australia antigen in 197 later known as HBs antigen- combined with viral hepatitis (3, 4) was a major breakthrough in the diagnosis of hepatitis B. HBs antigen appears several days to several weeks after contact with the virus and can persist for several months. Persistence of HBs antigen for more than 6 months serologically defines chronic HBV infection. VIKIA HBsAg enables the detection of HBsAg during acute infection or chronic carriage of the hepatitis B virus. PRINCIPLE VIKIA HBsAg is a qualitative test based on the association of monoclonal and polyclonal antibodies specific to HBsAg. This test uses the principle of lateral immunochromatography for the detection of circulating HBs antigen. It is used for the detection of the main subtypes ad and ay in serum, plasma, and whole. The test consists of a plastic device (see illustration on page 3) containing: 1. a chromatography membrane to which are fixed: - in the test region, a goat polyclonal anti-hbs antibody (test region "T") - in the control region, a monoclonal anti-biotin antibody (control region "C"). 2. A test strip impregnated with a conjugate consisting of: - a mixture of two monoclonal anti-hbs antibodies coupled to red-dyed polystyrene microspheres, - a BSA-biotinylated complex coupled to blue-dyed polystyrene microspheres. The sample is added to the sample well and migrates by capillarity along the membrane. If the sample contains HBs antigen, the HBs antigen forms an antigen-antibody complex with the antibodies specific to this virus present on the red-dyed polystyrene microspheres. The antigen-antibody complexes migrate along the membrane and bind to the anti-hbs antibodies forming complexes revealed by a red line in test region T of the membrane. To serve as a procedural control, a blue line will always appear in control region "C" if the test has been performed correctly. The BSA-biotinylated complex coupled to blue dyed polystyrene microspheres migrates along the membrane at the same time as the sample and binds to the antibiotin antibody forming a complex revealed by a blue line in control region C. Absence of this colored line invalidates the test. CONTENT OF THE KIT (25 TESTS) : 25 sealed pouches R1 Each pouch contains: - a ready-to-use test device (goat polyclonal anti-hbs antibody + mouse monoclonal anti-biotin antibody + polystyrene microspheres sensitized with mouse monoclonal anti-hbs antibody and a BSA-biotinylated complex). - a disposable specimen dropper. 1 Dropper bottle for whole 3ml Visual reading card 1 Package insert R2 MATERIAL REQUIRED BUT NOT PROVIDED - Timer. Ready-to-use. Stable 18 months after opening. Phosphate buffer ph g/l casein +.2 g/l sodium azide. Whole collection (fingerstick) - Lancets µl EDTA or lithium heparin capillary tubes. - Disposable pipette filler bulbs. biomérieux Brasil S.A. English - 1

2 VIKIA HBs Ag B - en - 26/11 WARNINGS AND PRECAUTIONS For in vitro diagnostic use only. For professional use only. This kit contains products of animal origin. Certified knowledge of the origin and/or sanitary state of the animals does not totally guarantee the absence of transmissible pathogenic agents. It is therefore recommended that these products be treated as potentially infectious, and handled observing the usual safety precautions (do not ingest or inhale). Do not use reagents past the expiry date indicated on the packaging. The test device should be stored in the sealed pouch containing the desiccant until use. The test device is a disposable; it should not be reused. All specimens should be considered infectious and handled following the recommended precautions (CLSI/NCCLS M29-A, Protection of Laboratory Workers from Instrument Biohazards and Infectious Disease Transmitted by Blood, Body Fluids, and Tissue; Approved Guideline Current revision). For additional information on handling precautions, refer to "Biosafety in Microbiological and Biomedical Laboratories CDC/NIH Latest edition", or the current regulations in the country of use. Do not mix reagents from different lots. Kit reagents contain sodium azide which can react with lead or copper plumbing to form explosive metal azides. If any liquid containing sodium azide is disposed of in the plumbing system, drains should be flushed with water to avoid build-up. STORAGE CONDITIONS AND STABILITY Store the kit at 4-3 C. DO NOT FREEZE. If stored according to the recommended conditions, all components are stable until the expiry date indicated on the packaging. Do not use after the expiry date. The test device should remain in the pouch until use. SPECIMEN COLLECTION AND PREPARATION 2. Whole collection by venipuncture Use whole collected by venipuncture on lithium heparin or EDTA. 3. Whole collection by fingerstick Use a capillary tube to collect from the fingertip. The sample should be used extemporaneously. Specimen stability Samples (serum and plasma) can be stored for 5 days at 2-8 C and 4 hours at C. If longer storage is required, freeze at - 25 ± 6 C. A study performed on frozen samples over a period of 2 months, showed that the quality of results is not affected. The venipuncture whole can be stored for 5 days at 2-8 C and 4 hours at C. Do not freeze whole samples. Whole collected by fingerstick should be tested immediately. INSTRUCTIONS FOR USE Allow the required reagents to come to room temperature before use. Serum or plasma samples 1. Remove the test device from the sealed pouch and use it as soon as possible. 2. Place the test device on a clean and level surface. 3. Using the dropper, transfer 3 drops of sample (approx. 75 µl) to the sample well (S) of the test device avoiding the formation of bubbles and then start the timer. See the illustration below. 4. After transferring the sample, wait for control line C to appear and possibly test line T. Read the test from the 15th minute onwards. Do not report a negative result before 3 minutes*. 3 drops of serum, plasma Specimen type and collection 1. Collection of serum, plasma Use serum or plasma collected in lithium heparin or EDTA. Serum and plasma should be stored separated from the pellet. The results obtained were not found to be influenced for icteric samples (bilirubin concentrations up to 5 µmol/l), for hemolyzed samples (hemoglobin concentrations up to 27 µmol/l) and for lipemic samples (up to 3 mg/ml). Do not inactivate samples. biomérieux Brasil S.A. English - 2

3 VIKIA HBs Ag B - en - 26/11 Whole sample (venipuncture) 1. Remove the test device from the sealed pouch and use it as soon as possible. 2. Place the test device on a clean and level surface. 3. Using the dropper, transfer three drops of whole (approx. 75 µl) to the sample well (S) of the test device. 4. Dispense one drop of buffer (approx. 4 µl), avoiding the formation of bubbles in the sample well (S). 5. Start the timer. See the illustration below. 6. After transferring the sample, wait for control line C to appear and possibly line T. Read the test from the 15th minute onwards. Do not report a negative result before 3 minutes*. 3 drops of venipuncture whole 1 drop of buffer Whole sample (finger-stick) 1. Remove the test device from the sealed pouch and use it as soon as possible. 2. Place the test device on a clean and level surface. 3. Collect approximately 75 µl of sample using the capillary tube. 4. Using the bulb, dispense in the sample well. 5. Dispense one drop of buffer (approx. 4 µl), avoiding the formation of bubbles in the sample well (S). 6. Start the timer. See the illustration below. 7. After transferring the sample, wait for control line C to appear and possibly test line T. Read the test from the 15th minute onwards. Do not report a negative result before 3 minutes*. Approx. 75 µl of fingerstick whole 1 drop of buffer * For some positive samples, the line in test region T may appear between 15 and 3 minutes. biomérieux Brasil S.A. English - 3

4 VIKIA HBs Ag B - en - 26/11 INTERPRETATION OF RESULTS POSITIVE: Two distinct lines appear: a blue line in the control region (C), and a red line in the test region (T). A pink to red line (T), even if it is very thin, indicates a positive result. NEGATIVE: A blue line appears in the control region (C). No line appears in test region T. INVALID: The control line fails to appear or no line appears in C and T ; insufficient specimen volume or incorrect procedural techniques are the most likely reasons. Review the procedure and repeat the test with a new test device. If the problem persists, discontinue using the kit and contact your local distributor. NOTE: The intensity of the red line in test region "T" may vary depending on the concentration of HBs antigen in the sample. However, the concentration of antigen in the sample cannot be determined by this qualitative test. For aid with reading, refer to the visual reading card. Interpretation of test results should be made taking into consideration the patient s history, and the results of any other tests performed. Positive Negative Invalid QUALITY CONTROL Internal procedural controls are included in the test. A blue colored line appearing in the control region (C) confirms sufficient specimen volume and correct procedural technique. If the control line does not appear, the test is invalid. Note It is the responsibility of the user to perform Quality Control in accordance with any local applicable regulations. LIMITATIONS OF THE METHOD DO NOT INTERPRET THE TEST AFTER MORE THAN 6 MINUTES. A negative HBsAg result does not allow infection by the hepatitis B virus to be excluded. The HBsAg serum concentration may in fact be below the analytical sensitivity of the reagent. The presence of a modified HBs antigen (variant) cannot be excluded; the antigen may, in this case, have been incorrectly recognized or not recognized by the antibodies in the reagent. The results of this test must be interpreted taking into account the overall clinical evaluation and the results of any other tests performed (Elisa and neutralization, HBV DNA, other serological markers, etc.). If both HBs antigen and anti-hbs antibodies are present, the quantity of antigen may be reduced, or negative in rare cases. This test has been validated for serum, plasma, and whole. It should not be used for other biological fluids such as saliva, CSF, or urine. This test should not be used with specimens collected post-mortem. Do not use serum pools. PERFORMANCE To take into account the different HBV genotypes, VIKIA HBsAg performance was studied as part of an international multicenter evaluation performed in West Africa (Burkina-Faso), South America (Brazil), and Asia (India and China). Plasma and serum sample status (EDTA) was established using the Elisa technique. VIKIA HBsAg performance: relative specificity and sensitivity on fresh plasma, serum and venous whole, was established in comparison with this Elisa technique. An equivalence study was performed for fresh plasma, venous whole, and capillary, on a limited number of paired samples. Furthermore, VIKIA HBsAg was evaluated using plasma or serum in comparison with another ICT test. biomérieux Brasil S.A English - 4

5 VIKIA HBs Ag B - en - 26/11 CLINICAL SPECIFICITY AND SENSITIVITY 18 paired plasma and venous whole samples, and 15 serum samples, (1528 negative and 422 positive) from different populations: donors, pregnant women, asymptomatic patients at testing centers, positive HBs antigen patients known to have chronic or acute hepatitis, were tested using the VIKIA HBsAg test. West Africa South America (Brazil) Asia Number of samples tested Sample status Negative Positive Asymptomatic patients and donors Pregnant women Positive HBs Ag patients Plasma ou serum number of samples per country A) PLASMA OR SERUM Reading at 3 minutes Elisa VIKIA HBsAg Positive Negative 4* * One sample with a positive status, that was found to be negative at 3 minutes, became positive between 3 and 6 minutes. Relative specificity of the VIKIA HBsAg test 99.8% [99.41% %] of 1528 negative plasma or serum samples. Relative sensitivity of the VIKIA HBsAg test Relative sensitivity at 15 and 3 minutes: 99.5% [97.55% %] of 422 positive plasma or serum samples. Relative sensitivity at 6 minutes: 99.29% [97.89% %] of 422 positive plasma or serum samples. B) VENOUS WHOLE BLOOD Reading at 3 minutes Elisa VIKIA HBsAg Positive Negative 4* * One sample with a positive status (same patient as for the Plasma study), that was found to be negative at 3 minutes, became positive between 3 and 6 minutes. Relative specificity of the VIKIA HBsAg test 99.79% [99.37% %] of 1428 venous whole samples. Relative sensitivity of the VIKIA HBsAg test Relative sensitivity at 15 and 3 minutes: 98.92% [97.22% %] of 372 positive venous whole samples. Relative sensitivity at 6 minutes: 99.19% [97.61% %] of 372 positive venous whole samples. biomérieux Brasil S.A English - 5

6 VIKIA HBs Ag B - en - 26/11 ANALYTICAL SENSITIVITY The analytical sensitivity was determined using the WHO standard (Second International Standard for HBsAg, subtype adw2, genotype A). It is less than or equal to 2 IU/ml (3.8 ng/ml). COMPARISON WITH ANOTHER ICT TEST This comparison is performed using plasma or serum samples only. a) samples with a negative status VIKIA HBsAg Other ICT Positive test Negative Agreement : 99,8% 3 samples with a negative status were found to be positive with the VIKIA test. b) samples with a positive status VIKIA HBsAg Other Positive 412 1* 413 Negative * A sample that was negative with the VIKIA test at 3 minutes became positive between 3 and 6 minutes Agreement: Agreement at 15 and 3 minutes: 98.34% Agreement at 6 minutes: 98.58% 6 samples with a positive status that were detected by the VIKIA test, were not detected by the other ICT test. EQUIVALENCE STUDY OF THE DIFFERENT TYPES OF SPECIMENS This study was performed using VIKIA HBsAg on 126 samples: 95 samples with a negative status and 31 paired samples with a positive status: fresh plasma, venous whole and capillary whole. VIKIA HBsAg Sample status Number of samples tested Plasma Venous whole Capillary whole Negative Positive 31 32* 32* 32* 126 *One sample with a negative status was found to be positive with the VIKIA HBsAg test whatever the type of specimen used. No discrepancies were observed between the different types of specimens. biomérieux Brasil S.A English - 6

7 VIKIA HBs Ag B - en - 26/11 WASTE DISPOSAL Dispose of used or unused reagents as well as any other contaminated disposable materials following procedures for infectious or potentially infectious products. It is the responsibility of each laboratory to handle waste and effluents produced according to their nature and degree of hazardousness and to treat and dispose of them (or have them treated and disposed of) in accordance with any applicable regulations. LITERATURE REFERENCES 1. MILICH D.R., Immune response to the hepatitis B virus : infection, animal models, vaccination, VIRAL HEPATITIS, 1997, 3, HOLLINGER F.B., Hepatitis B virus, in Fields Virology, Third Edition, Lippincott-Raven Publishers, Phildelphia, 1996, BLUMBERG B.S., ALTER H.J., VISNICH S. JAMA, A "New" Antigen in Leukemia Sera, 1965, 191, PRINCE A.M., An antigen detected in during the incubation period of serum hepatitis, Proc Natl Acad Sci USA, 1968, 6, INDEX OF SYMBOLS Symbol Meaning Catalogue number In Vitro Diagnostic Medical Device Manufacturer Temperature limitation Use by Batch code Consult Instructions for Use Contains sufficient for "n" tests Do not reuse WARRANTY biomérieux disclaims all warranties, express or implied, including any implied warranties of MERCHANTABILITY AND FITNESS FOR A PARTICULAR USE. biomérieux shall not be liable for any incidental or consequential damages. IN NO EVENT SHALL BIOMERIEUX S LIABLITY TO CUSTOMER UNDER ANY CLAIM EXCEED A REFUND OF THE AMOUNT PAID TO BIOMERIEUX FOR THE PRODUCT OR SERVICE WHICH IS THE SUBJECT OF THE CLAIM. biomérieux Brasil S.A. Estrada Do Mapua 491 Taquara Jacarepagua CEP RIO DE JANEIRO R.J. Brasil Tél. 33 () Fax 33 () Printed in France biomérieux, the blue logo and VIKIA are used, pending and/or registered trademarks belonging to biomérieux SA or one of its subsidiaries.