HIV Ab&Ag
The kit is a solid phase enzyme immunoassay for the determination of antibodies to all subtypes of HIV-1 and HIV-2 and HIV-1 antigen (p24) in human serum or plasma. The kit may be used for the screening of blood units and the follow up of HIVinfected patients. This kit is intended exclusively for In vitro diagnostic use in an authorized clinical laboratory and the test has to be carried out by specifically trained health-care professional personnel.
Epidemiological evidence indicates that an infectious agent transmitted through intimate contact, intravenous drug use or use of infected blood or blood products leads to Acquired Immunodeficiency Syndrome (AIDS).
This disease affects T-cell mediated immunity, resulting in severe lymphopenia and a reduced subpopulation of helper T-lymphocytes. Destruction of this T-lymphocyte population by the virus causes an immune deficiency, resulting in a reduced or deficient response to subsequent infections.
Consequently, infections become more severe and may cause death. At present, there is no successful treatment for AIDS.
The etiological agent has been identified as a retrovirus, human immunodeficiency virus type 1 (HIV-1).
A closely related, but distinct type of immunodeficiency virus, designated HIV-2, has also been isolated. This virus causes a disease that is indistinguishable from AIDS.
Serological cross-reactivity between HIV-1 and HIV-2 has been shown to be highly variable from sample to sample.
This variability requires the inclusion of antigens to both HIV-1 and HIV-2 for the screening of antibodies to HIV-1 and HIV-2.
The presence of anti-HIV-1 and/or anti-HIV-2 and/or HIV p24 antigen in the blood indicates potential infection with HIV-1 and/or HIV-2 and consequently this blood should not be used for transfusion or for manufacture of injectable products.
Synthetic peptides representing immunodominant epitopes of HIV-1 and HIV-2 together with a monoclonal antibody to p24 HIV-1 antigen are coated onto wells of a microplate.
The peptides and the antibody have been carefully selected to ensure the screening of antibody and p24 antigen to all HIV-1 subtypes, including subtype O and HIV-2. Serum or plasma samples are added to these wells and, if antibodies specific to HIV- 1 and/or HIV-2 (IgG, IgM or IgA) are present in the sample, they will form stable complexes with the HIV peptide antigens in the well. In case HIV-1 p24 is present in the sample, the antigen will be captured by the specific monoclonal antibody.
Antigen-antibody complexes are then identified through the successive addition of: (1) biotinylated peptides, a biotinylated monoclonal antibody to HIV-1 p24, and; (2) horseradish peroxidase HRP Streptavidin conjugate.
The hydrolytic activity of horseradish peroxidase allows for the quantification of these antibody-antigen complexes. Peroxidase substrate solution is then added.
During incubation, a blue color will develop in proportion to the amount of anti-HIV-1/2 antibodies or HIV-1 p24 antigen bound to the well, thus establishing their presence or absence in the sample. Wells containing samples negative for anti-HIV antibody and/or p24 antigen remain colorless.
A stop solution is added to each well and the resulting yellow color is read on a microplate reader at 450 nm.
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- Alizon, M., Sonigo, P., Barré-Sinoussi, F., Chermann, J.-C., Tiollais, P., Montagnier, L. and Wain-Hobson, S., 1984. Molecular Cloning of Lymphadenopathy-Associated Virus. Nature 312: 757-760.
- Hahn, B.N., Shaw, G.M., Arya, S.K., Popovic, M., Gallo, R.C. and Wong-Staal, F., 1984. Molecular Cloning and Characterization of the HTLV-III Virus Associated with AIDS. Nature 312: 166-169.
- Luciw, P.A., Potter, S.J., Steimer, K., Dina, D. and Levy, J.A., 1984. Molecular Cloning of AIDS-Associated Retrovirus. Nature 312: 760-763.
- Popovic, M., Sarngadharan, M.G., Read, E. and Gallo, R.C., 1984. Detection, Isolation, and Continuous Production of Cytopathic Retroviruses (HTLV-III) from Patients with AIDS and Pre-AIDS. Science 224: 497-500.
- Sarngadharan, M.G., Popovic, M., Bruch, L., Schüpbach, J. and Gallo, R.C., 1984. Antibodies Reactive with Human T-Lymphotropic Retroviruses (HTLV-III) in the Serum of Patients with AIDS. Science 224: 506-508.
- Vézinet-Brun, F., Barré-Sinoussi, F., Saimot, A.G., Christol, D., Montagnier, L., Rouzioux, C., Klatzmann, D., Rozenbaum, W., Gluckmann, J.C. and Chermann, J.-C., 1984. Detection of IgG Antibodies to Lymphadenopathy-Associated Virus in Patients with AIDS or Lymphadenopathy Syndrome. Lancet: 1253-1256, June 9.
- Spire, B., Montagnier, L., Barré-Sinoussi, F. and Chermann, J.-C., 1984. Inactivation of Lymphadenopathy Associated Virus by Chemical Disinfectants. Lancet: 899-901, Oct. 20.
- Sehulster, L.M., Hollinger, F.B., Dreesman, G.R. and Melnick, J.L., 1981. Immunological and Biophysical Alteration of Hepatitis B Virus Antigens by Sodium Hypochlorite Disinfection. App. and Envir. Microbiol. 42: 762-767.
- Barre-Sinoussi, F., Chermann, J.C., Rey, F., Nugeyre, M.T., Chamaret, S., Gruest, J., Dauguet, C., Axler-Blin, C., Vezinet- Brun, F., Rouzioux, C., Rozenbrum, W. et Montagnier, L. 1983. Isolation of a T-lymphotropic retrovirus from a patient at risk for acquired immune deficiency syndrome (AIDS). Science 220: 868-871.
- Gallo, R.C., Salahuddin, S.Z., Popovic, M., Shearer, G.M., Kaplan, M., Haynes, B.F., Palker, T.J., Redfield, R., Oleske, J., Safai, B., White, G., Foster, P. and Markham, P.D. 1984. Frequent detection and isolation of cytopathic retroviruses (HTLV-III) from patients with AIDS and at risk for AIDS. Science 224:500-503.