TRUSTwell HIV Ag-Ab 4th Gen ELISA

TEST PRINCIPLE  

The TRUSTwell HIV Ag-Ab ELISA Kit is a solid phase enzyme linked immunosorbent assay based on the principle of the double antibody/antigen sandwich technique for the detection of HIV-1 P24 antigen and various antibodies against HIV-1 and/or HIV-2 antigen in human serum or plasma. The TRUSTwell HIV Ag-Ab ELISA Kit is composed by two systems: 1) Solid microwells pre-coated with Recombinant HIV-1 and HIV-2 antigens and anti-P24 antibody 2) Liquid conjugates composed of a. Recombinant HIV-1 and HIV-2 antigens conjugated with horse radish peroxidase (HRP-HIV 1+2 conjugates) and avidin conjugated with HRP (HRP-avidin conjugates) b. Anti-P24 antibody conjugated with biotin (Biotinylated P24 Ab)

During the assay, the test specimen is first incubated with the coated microwells and followed by incubation with biotinylated P24 Ab. The anti-HIV-1 and anti-HIV-2 antibodies, if present in the specimen, bind to the antigens coated on the microwell surface. If P24 present in the specimen, binds to the anti-P24 antibody on the microwell surface and biotinylated P24 Ab in the solution. In the next incubation with the HRP-HIV 1+2 and HRP-avidin conjugates, the above bounded anti-HIV-1 and anti-HIV-2 antibodies and bounded P24 antigen reactive to the HRP conjugates, forming HRP-complex. Unbounded conjugates are then removed by washing. The presence of the complexed conjugates is shown by a blue color upon additional incubation with TMB substrate. The reaction is stopped with Stop Solution and absorbance are read using a spectrophotometer at 450/620-690nm.

STORAGE AND STABILITY

All reagents except the concentrated wash buffer are ready to use as supplied. Return all reagents requiring refrigeration immediately after use. Reseal the microwells after removing the desired number of wells. Ensure that the reagents are brought to room temperature before opening. All the reagents are stable through the expiration date printed on the label if not opened. Do not freeze the kit or expose the kit over 8’C