This HIV (1+2+O) Ag/Ab ELISA kit is a two-step incubation, “sandwich” enzyme immunoassay, which uses polystyrene microwell strips pre-coated with recombinant HIV antigens (recombinant HIV-1gp41, gp120, antigens specific for the detection of HIV-1 group O, and recombinant HIV-2 gp-36), and anti-HIV (p24) antibodies.
As a first step, biotinylated anti-HIV (p24) antibodies together with the patient’s serum or plasma sample are added into the wells. During incubation, the specific HIV1/2 antibodies if present in sample, will be captured inside the wells.
Simultaneously, if HIV p24 antigen is present in sample, it will also be captured as a double antibody “sandwich” complex comprising of the coated antibodies-p24-biotinylated antibodies. The microwells are then washed to remove unbound serum proteins.
The detection of the captured HIV p24 antigen-biotinylated antibody complex or HIV 1/2 antibodies is achieved during the second incubation step by adding of the enzyme Horseradish Peroxidase (HRP) which has been conjugated to second HIV 1+2 recombinant antigens and to avidin.
p24 detection:
When p24 has been captured inside the wells, avidin will react with the biotin and attach HRP to the Ab-p24-Ab complex.
HIV 1/2 antibody detection:
When HIV 1/2 antibodies have been captured inside the wells, the HRP-conjugated antigens will bind to the captured antibodies forming Ag-Ab-Ag(HRP) “sandwich” immunocomplex.
The microwells are washed to remove unbound conjugate, and Chromogen solutions are added to the wells.
In wells containing the Ag-Ab-Ag (HRP) and/or Ab-p24-Ab(HRP) “sandwich” immunocomplexes, the colorless Chromogens are hydrolyzed by the bound HRP to a blue colored product. The blue color turns yellow after stopping the reaction with sulfuric acid. The amount of color intensity can be measured and is proportional to the amount of antibodies or p24 captured in the wells, and to the sample respectively. Wells containing samples negative for anti-HIV 1/2 or p24 remain colorless.
96 Tests
l MICROWELL PLATE 1plate
Blank microwell strips fixed on white strip holder. The plate is sealed in aluminium pouch with desiccant.
8×12/12×8-well strips wells per plate. Each well contains recombinant HIV 1/2 antigens and anti-p24 antibodies. The microwell strips can be broken to be used separately. Place unused wells or strips in the plastic sealable storage bag together with the desiccant and return to 2~8℃.
l NEGATIVE CONTROL 1vial
Yellowish liquid filled in a vial with green screw cap
1ml per vial.
Protein-stabilized buffer tested non-reactive for HIV 1/2.
Preservatives: 0.1% ProClin 300.
Ready to use as supplied.
Once open, stable for one month at 2-8℃.
l ANTIBODY POSITIVE CONTROL-1 (HIV 1) 1vial
Red-colored liquid filled in a vial with red screw cap
1ml per vial
Antibodies to HIV 1 diluted in protein-stabilized buffer
Preservatives: 0.1% ProClin 300.
Ready to use as supplied.
Once open, stable for one month at 2-8℃
l ANTIBODY POSITIVE CONTROL-2 (HIV 2) 1vial
Red-colored liquid filled in a vial with yellow screw cap.
1ml per vial
Antibodies to HIV 2 diluted in protein-stabilized buffer
Preservatives: 0.1% ProClin 300.
Ready to use as supplied.
Once open, stable for one month at 2-8℃.
l ANTIGEN POSITIVE CONTROL 1vial
Red-colored liquid filled in a vial with blue screw cap.
