Optimization Of An Elisa Assay For Detecting Hiv-I Mper Antibodies In HIV-I Discordant Couples

Abstract

ELISA techniques have been of great use in antigen and antibody detection. In this study, we have optimized an in-house indirect ELISA technique for antigenicity assays involving HIV-I discordant couples.  We investigated the ability of the HIV-I Q?MPERN protein to be used in detecting antibody responses in HIV-I seropositive and highly exposed but seronegative individuals. This study was aimed at developing and enhancing an Indirect ELISA Assay technique. Our group had engineered the hybrid phage HIV-I Q?MPERN. Optimal requirements for ELISA assays, which include the concentration of the coating antigen, labeled antibody concentrations, temperatures and incubation times were determined. To confirm the efficacy of our assay technique, ethical approval was obtained, informed consent sought and questionnaire were administered. Serum samples were collected from 24 HIV-I discordant couples, analyzed and results were read using Thermofisher multiscan ELISA reader. The 31-40 years age bracket had the most infection HIV-I infection while the 51-60 years age bracket were least infected by HIV-I. Older men had more HIV-I infection than older women while younger women were more infected than younger men. Forty two percent of men and 58% of women were positive for HIV-I among the discordant couples. Optimal conditions were considered in performing the ELISA assay. The IgG antibody response obtained in this study was strong for both the HIV-I seropositive and seronegative couples. This study has confirmed the efficacy of our optimized in-house ELISA assay at detecting antibody responses associated with the HIV-I MPER. The assay was found to be efficacious and cost effective.