Human Immunodeficiency Virus 1 and 2 Antigen and Antibody Cut-Off ELISA 96 Wells Dialab

The human immunodeficiency viruses (HIVs) type 1 and type 2 are the etiological agents of the acquired immunodeficiency syndrome (AIDS). HIV has been isolated from patients with AIDS, AIDS related complex (ARC) and from healthy individuals at high risk for AIDS. Infection with HIV is followed by an acute flu-like illness. This phase may remain unnoticed and the relationship to HIV infection may not be clear in many cases. The acute phase is typically followed by an asymptomatic carrier state, which progresses to clinical AIDS in about 50% of infected individuals within 10 years after seroconversion. Serological evidence of infection with HIV may be obtained by testing for the presence of HIV antigens or antibodies in the serum of individuals with suspected HIV infection. Antigens can generally only be detected during both acute and symptomatic phases. The antibodies to HIV-1 and/or HIV-2 can be detected virtually throughout the whole; infection period, starting at or shortly after, the acute phase and lasting till the end stage of AIDS. Apart from sexual transmission, the principle route of infection with HIV is blood transfusion. All donations of blood or plasma should be tested due to the risk of HIV transmission through contaminated blood. This is a 4th generation HIV ELISA that simultaneously detects HIV antigens (gp41, gp120, gp36) and antibodies directed at HIV p24.

This assay is intended for the qualitative determination of antigens and/or antibodies to HIV type 1 and/or type 2 in serum or plasma. The test wells are coated with recombinant HIV1/2 antigens and anti-p24 antibodies. During the first incubation step patient sample and Biotin-conjugated anti-HIV (p24) are added to the well. If HIV-1/2 antibodies are present they will be bound to the well, if HIV p24 antigen is present is will be capture in th well as a double antibody sandwich complex. Free and bound are separated by washing and then horseradish peroxidase (HRP) conjugated anti-human IgG which bind the immobilised antigen- antibody complex. Unbound conjugate is removed by washing and then enzyme substrate added which in the presence of the conjugate hydrolyses to form a blue colour. Addition of stop solution forms a yellow end-product which is photometrically measured at 450nm. The amount of colour is proportional to the concentration of HIV antibodies and/or p24 antigen in the patient sample. After validation checks a Cut-Off Value is calculated.
Negative: < 1 Cut-off Value
Positive: ≥ 1 Cut-off Value
Borderline 0.9 - 1.1 Cut-off Value (Re-test to confirm)

Coated Microtiter wells : 12x8 well strips
Antibody Positive Control HIV1: 1ml
Antibody Positive Control HIV2: 1ml
Antigen Positive Control: 1ml
Negative Control: 1ml
Enzyme Conjugate : 12ml
Biotin Conjugate :3ml
Substrate Solution A : 1x6ml
Substrate Solution B : 1x6ml
Stop Solution: 1x6ml
Wash buffer (20X); 50ml
Sample volume: 100µl human serum or plasma
Incubation time: 2h (60 min, 30min, 30min all at 37°C)
Store at 2-8°C.