HIV Ag-Ab Test PDF

36 Determine™ HIV–1/2 Ag/Ab Combo INTENDED USE Alere Determine™ HIV–1/2 Ag/Ab Combo is an in vitro, visually read, qualitative immunoassay for the simultaneous detection of Human Immunodeficiency Virus Type 1 (HIV-1) p24 antigen (Ag) and antibodies (Ab) to HIV Type 1 and Ty...

36 Determine™ HIV–1/2 Ag/Ab Combo INTENDED USE Alere Determine™ HIV–1/2 Ag/Ab Combo is an in vitro, visually read, qualitative immunoassay for the simultaneous detection of Human Immunodeficiency Virus Type 1 (HIV-1) p24 antigen (Ag) and antibodies (Ab) to HIV Type 1 and Type 2 (HIV-1 and HIV-2) in human serum, plasma, capillary (fingerstick) whole blood or venipuncture (venous) whole blood. It is intended for use as a point-of-care test to aid in the diagnosis of infection with HIV-1 and HIV-2, including an acute HIV-1 infection, and may distinguish acute HIV-1 infection from established HIV-1 infection when the specimen is positive for HIV-1 p24 antigen and negative for anti-HIV-1 and anti-HIV-2 antibodies. The test is suitable for use in multi-test algorithms designed for the statistical validation of rapid HIV test results. When multiple rapid HIV test are available, this test can be used in appropriate multi-test algorithms. Alere Determine™ HIV-1/2 Ag/Ab Combo is not intended for newborn screening or for use with cord blood specimens or specimens from individuals less than 12 years of age. Alere Determine™ HIV-1/2 Ag/Ab Combo is not intended for use in screening blood, plasma, cell, or tissue donors. CLIA Complexity: Moderate SUMMARY AND PRINCIPLE OF TEST Alere Determine™ HIV–1/2 Ag/Ab Combo is an immunochromatographic test for the simultaneous and separate qualitative detection of free HIV-1 p24 antigen and antibodies to HIV-1 and HIV-2. The test device is a laminated strip that consists of a Sample Pad containing monoclonal biotinylated anti-HIV-1 p24 antibody, a Conjugate Pad containing monoclonal anti-HIV-1 p24 antibody-colloidal selenium and HIV-1 and HIV-2 recombinant antigen-colloidal selenium, and a nitrocellulose membrane with an immobilized mixture of recombinant and synthetic peptide HIV-1 and HIV-2 antigens in the Lower Test Area, immobilized streptavidin in the Upper Test Area, and an immobilized mixture ofanti-HIV-1 antibodies, HIV-1/2 antigens, and HIV-1 p24 recombinant antigen and anti-HIV-1 p24 monoclonal antibody in the Control Area. SP = Sample Pad; CP = Conjugate Pad; LTA = Lower Test Area; UTA = Upper Test Area; CA = Control Area A specimen (venipuncture or capillary whole blood, serum, or plasma) is applied to the Sample Pad (followed by Chase Buffer for venipuncture or fingerstick whole blood specimens) and migrates by capillary action through the Conjugate Pad and then through the nitrocellulose membrane. If HIV - 1 p24 antigen is present in the specimen; it binds with the monoclonal biotinylated anti-HIV-1 p24 antibody from the Sample Pad and then with monoclonal anti-HIV-1 p24 antibody-colloidal selenium from the Conjugate Pad to form a complex (biotinylated antibody-antigen-colloidal selenium-antibody). This complex migrates through the solid phase by capillary action until it is captured by immobilized streptavidin at the Upper Test Area (labeled “Ag”) where it forms a single pink/red “Ag” line. If HIV-1 p24 antigen is not present in t h e specimen or is below the limit of detection of the test, no pink/red Ag line is formed. NOTE: The monoclonal biotinylated anti-HIV-1 p24 antibody used in this assay does not cross react with HIV-2 p26 antigen.  37 If antibodies to HIV-1 and/or HIV-2 are present in the specimen, the antibodies bind to recombinant gp41 (HIV- 1) and gp36 (HIV-2) antigen-colloidal selenium conjugates from the Conjugate Pad. The complex migrates through the solid phase by capillary action until it is captured by immobilized HIV-1 and HIV-2 synthetic peptide antigens and recombinant gp41 antigen at the Lower Test Area (labeled “Ab”) and forms a single pink/red “Ab” line. If antibodies to HIV-1 and/or HIV-2 are absent or are below the detection limit of detection of the test, no pink/red Ab line is formed. HIV 1 p24 recombinant antigen and anti-HIV-1 p24 monoclonal antibody is incorporated in the nitrocellulose membrane. For a testresult to be valid there must be a visible pink/red Control line. During the testing procedure the colloidal selenium conjugates released from the Conjugate Pad will be captured by the antibodies and antigens immobilized in the Control Area and form a pink/red Control line for samples that are either positive or negative. COMPONENTS OF ALERE DETERMINE™ HIV-1/2 AG/AB COMBO: Materials Provided Alere Determine™ HIV-1/2 Ag/Ab Combo Cards. Each Card consists of 5 or 10 Test Units which can be separated from each other by tearing along the perforated lines. Each Test Unit has a cover that is to be removed for sample application and visualization of test results. Desiccant Package Chase Buffer: Containing sodium chloride, disodium hydrogen phosphate, and Nipasept as a preservative. Quick Reference Guide Package Insert Subject Information Notices: 25 in the 25 Test Units kit, and 100 in the 100 Test Units kit. Customer Letter Disposable Capillary Tubes: For collection and transfer of fingerstick samples. Disposable Workstations: 25 in the 25 Test Units kit, and 100 in the 100 Test Units kit. Internal Control Features Test contains built-in control features. The manufacturer’s recommendation for daily quality control is to document these controls for the first sample tested each day. Quality Control log sheets are available from Technical Support. No additional external quality control is required. ◼ Internal Positive Procedural Control: A pink/red line in the Control area is considered an internal positive procedural control. If the test has been performed correctly and the Reaction Unit is working properly, this indicator will appear. ◼ Internal Negative Procedural Control: A clear background is considered an internal negative procedural control. If the test has been performed correctly and the Reaction Unit is working properly, the background will clear to give a discernable result.  38 MATERIALS REQUIRED, BUT NOT PROVIDED Clock, watch, or other timing device Precision pipette capable of delivering 50μL of sample with disposable tips, to be used in lieu of the Disposable Capillary Tubes supplied with the kit (for other than fingerstick whole blood specimens) Disposable gloves Sterile gauze (for fingerstick whole blood specimens) Antiseptic wipes Biohazard disposal container Collection devices for specimens (other than fingerstick whole blood specimens) TEST PROCEDURE Kit Component Preparation Remove the desired numbers of test units from the 5- or 10-Test Unit Card by bending and tearing at the perforation. NOTE: Removal of the test units should start from the right side of the Card to preserve thelot number which appears on the left side of the Card. Lay the Test Unit flat in the workstation and remove the protective foil cover from each TestUnit. The test should be initiated within 2 hours after removing the protective foil cover from each Test Unit. NOTE: Use of the workstation is optional. If the workstation is not used, place the Test Unit on a flat surface. For serum or plasma samples: 1. Using a micropipette, apply 50 μL of sample to the Sample Pad (marked by the arrow symbol). Donot add Chase Buffer when using serum or plasma specimens. 2. Read the test result between 20 and 30 minutes after the addition of the Sample. Do not read test results after 30 minutes. NOTE: Discard the used pipette tips, Capillary Tube, Test Units and any other test materials into a biohazard wastecontainer. INTERPRETATION AND REPORTING 8OF TEST RESULTS ANTIBODY REACTIVE (Two Lines - Control Line and Ab Line) A pink/red Control line appears in the Control Area AND a pink/red Ab line appears in theLower Test Area of the Test Unit. The intensity of the Ab and Control lines may vary. Anyvisible pink/red color in both the Control and Lower Test Areas, regardless of intensity, is considered REACTIVE. A Reactive test result means that HIV-1 and/or HIV-2 antibodies have been detected in the specimen. The test result is interpreted as PRELIMINARY REACTIVE. Report at as PRELIMINARY REACTIVE for HIV-1 &/or HIV-2 antibodies ANTIGEN (HIV-1p24) REACTIVE (Two Lines - Control Line and Ag Line) A pink/red Control line appears in the Control Area AND a pink/red Ag line appears in the UpperTest Area of the Test Unit. The intensity of the Ag and Control lines may vary. Any visible pink/red color in both the Control and Upper Test Areas, regardless of intensity, is considered REACTIVE. A Reactive test result means that HIV-1 p24 antigen has been detected in the specimen. The test result is interpreted as PRELIMINARY REACTIVE for HIV-1 p24 antigen. Report as PRELIMINARY REACTIVE for HIV-1 p24 antigen NOTE: A test result that is PRELIMINARY POSITIVE for HIV-1 p24 antigen in the absence of reactivity for HIV-1 or HIV-2 antibodies may indicate an acute HIV-1 infection in the test subject. In this case the acute HIV-1 infection is distinguished from an established HIV-1 infection in which antibodies to HIV-1are present.  39 ANTIBODY REACTIVE AND ANTIGEN (HIV-1 p24) REACTIVE (Three Lines - Control, Ab and Ag Lines) A pink/red Control line appears in the Control Area AND a pink/red Ab line appears in the Lower Test Area AND a pink/red Ag line appear in the Upper Test Area of the Test Unit. The intensityof the Ab, Ag and Control lines may vary. Any visible pink/red color in the Control Area, the Lower Test Area and the Upper Test Area, regardless of intensity, is considered REACTIVE. T The test result is interpreted as PRELIMINARY REACTIVE for HIV-1 and/or HIV-2 antibodies and HIV-1 p24 antigen. Report as PRELIMINARY REACTIVE for HIV-1 &/0r HIV-2 and HIV-1 p24 antigen NONREACTIVE (One Line – Control Line) A pink/red Control line appears in the Control Area of the Test Unit, and no pink/red Ab or Ag line appears in the Lower Test Area and the Upper Test Area of the Test Unit, respectively. A NONREACTIVE test result means that HIV-1 or HIV-2 antibodies and HIV-1 p24 antigen were not detected in the specimen. Report as NONREACTIVE INVALID (No Control Line) If there is no pink/red Control line in the Control Area of the Test Unit, even if a pink/red line appears in the Lower Test Area or the Upper Test Area of the Test Unit, the result is INVALIDand the test should be repeated. Report as INVALID: MUST REPREAT RESTRICTIONS This assay has not been evaluated for newborn screening or for use with cord blood specimens or for use with specimens from individuals less than 12 years of age. WARNINGS - For In Vitro Diagnostic Use 1. Read the Package Insert completely before using this assay. Follow the instructions carefully as not doing so may result in inaccurate test results. 2. Use of this test kit with specimen types other than those specifically approved for use with this device may produce inaccurate test results. 3. This test should be performed at 15 to 30°C (59 to 86°F). If stored refrigerated, ensure that the Test Units arebrought to operating temperature before performing testing. 4. Do not open or remove the protective foil cover from the Test Unit until just prior to use (Test Units should be usedwithin 2 hours of removing the protective foil). 5. Do not use kit contents beyond labeled expiration date. 6. Ensure that the test subject’s finger is completely dry before obtaining a fingerstick sample. 7. Ensure that a sample or Control is applied to the Sample Pad. Failure to apply a sample may give a false negative test result. 8. Read test results in a well-lit area. 9. Reading test results for serum or plasma specimens earlier than 20 minutes or later than 30 minutes after additionof the serum or plasma specimen may yield erroneous results. Reading test results for capillary (fingerstick) or venous (venipuncture) whole blood specimens earlier than 20 minutes or later than 30 minutes after addition of the Chase Buffer may yield erroneous results. 10. Specimens from individuals infected with HIV-1 and/or HIV-2 who are receiving “Highly Active Antiretroviral Therapy” (HAART) may produce false negative test results. 11. Specimens from individuals with Toxoplasma IgG, human anti-mouse antibodies, rheumatoid factor, elevated triglycerides (above 600 mg/dL), herpes simplex virus infection, and hospitalized and cancer patients may give false positive test results.  40 PRECAUTIONS 1. Safety Precautions Handle the samples, material contacting samples, and kit controls as if capable of transmitting infection. Wear protective clothing such as laboratory coats, disposable gloves, and eye protection when handling patient samples. Do not eat, drink, smoke, apply cosmetics, or handle contact lenses in areas where samples and kit reagent materials are handled. Avoid any contact between hands, eyes, or mouth during sample collection and testing. Decontaminate and dispose of all specimens, reagents, disposable workstations, and other potentially contaminated materials in a biohazard waste container in accordance with local regulations. Lancets should be placed in a puncture-resistant container prior to disposal. The recommended method of disposal of biohazard waste is autoclaving for a minimum of 1 hour at 121°C. Disposable materials may incinerated. Liquid wastes may be mixed with appropriate chemical disinfectants. A freshly prepared solution of 10% bleach (0.5% solution of sodium hypochlorite) is recommended. Allow 60 minutes for effective decontamination. NOTE: Do not autoclave solutions that contain bleach. The workstations are for singleuse only. The used workstation and Test Unit should be regarded as potentially infectious material. They should be disposed of together, without trying to remove the Test Unit from the workstation, in a biohazard waste container as indicated above. Clean and disinfect all spills of specimens or reagents using 10% bleach or other appropriate disinfectant.The bleach solution should be made fresh every day. For additional information refer to: Centers for Disease Control and Prevention: Updated U.S. Public Health Service Guidelines for the Management of Occupational Exposures to HBV, HCV, and HIV and Recommendation for Post-exposure Prophylaxis.7 2. Handling Precautions If Desiccant Package is missing, DO NOT USE. Discard Test Cards (all Test Units) and use a new TestCard. Do not use any Test Units from Test Cards if the pouch has been perforated. Each Test Unit, lancet and Disposable Capillary Tube for collection and transfer of fingerstick samples is for single use only. Do not use kit components beyond the expiration date printed on the label. Always check expiration date prior to testing. Adequate lighting is required to read a test result.