CLINICAL USE
Conventional immunoassays for the detection of antibodies against HIV may result inefficient in detecting primary infections since seroconversion occurs after the 3rd-4th week post-infection.
Combining simplicity, sensitivity, and precision in only one test, 4th generation assays simultaneously detect HIV antigens and antibodies and allow the early detection of the infection.

PRINCIPLE
The wells are coated with HIV-1 and HIV-2 recombinant proteins and synthetic peptides and anti-p24 monoclonal antibodies. The sample is incubated in the wells. If p24 and/or HIV-1 or HIV-2 antibodies are present in the sample, these will bind to the antigens and/or antibodies in the wells. The unbound material is removed by washing. Next, Conjugate 1 is added, which contains antibodies and antigens marked with biotin that will bind to the antigens/antibodies present in the sample. Then, Conjugate 2 is added (peroxidase-conjugated streptavidin), which will bind to Conjugate 1. The unbound conjugate is removed by washing.
Then, a solution of tetramethylbenzidine (TMB) and hydrogen peroxide is added. Reactive samples develop a blue color, which turns into yellow when the reaction is stopped with sulfuric acid (Stopper).

ADVANTAGES
• Simultaneous detection of specific HIV antigen & antibodies
• Two determinations in one
• Breakable wells
• Sample Addition Monitoring (SAM)
• Colored reagents for Process Control Monitoring (PCM)
• Biotin-Streptavidin-Peroxidase system
• Anti-p24 humanized antibody
• High contrast between positive and negative samples

PROVIDED REAGENTS
• Coated microtitration plate: 96 wells microtitration plate coated with HIV-1 and HIV-2 recombinant proteins and synthetic peptides and human anti-p24 monoclonal antibodies.
• Sample diluent: 0.02 M Tris Buffer containing bovine proteins, sodium chloride, and tensioactive agent pH 7.2, blue color.
• Conjugate 1: Ready-to-use solution containing HIV-1 and HIV-2 recombinant antigens and synthetic peptides and anti-p24 monoclonal humanized antibodies biotinylated in 10 mM phosphate buffer with bovine proteins, sodium chloride, and tensioactive agent pH 7.2.
• Concentrated Conjugate 2: Streptavidin conjugated to peroxidase, orange color.
• Conjugate 2 diluent: 10 mM Phosphate Buffer with bovine proteins, sodium chloride, and tensioactive agent pH 7.2.
• TMB: 36 mM Tetramethylbenzidine solution in 100% dimethyl sulfoxide (DMSO), (100x).
• TMB diluent: 40 mM Citrate buffer and 1.27 mM hydrogen peroxide, pH 4.3.
• Stopper: 1 M Sulfuric acid.
• Concentrated Wash Buffer: 250 mM Phosphate buffer, 3.45 M sodium chloride, and tensioactive agent, (25x), pH 6.4.
• Positive Control: Inactivated human serum with anti-HIV antibodies, red color.
• p24 Positive Control: HIV-1 p24 antigen solution in 10 mM phosphate buffer with bovine proteins, sodium chloride, and tensioactive agent, pH 7.2.
• Negative Control: Non reactive, inactivated, normal human serum, yellow color.

WIENER LAB. PROVIDES
Cat. N°:  1723451
Kits for 96 tests