Study of double-antibody sandwiched ELISA method to detect HIV-1 p24 antigen

Objective To establish a sensitive and specific method for the detection of HIV-1 p24 antigen.Methods Monoclonal and polyclonal antibodies were obtained from immunity mice and rabbits using purified HIV-1 p24 antigen expressed by genetic engineering.Antibodies were conjugated with HRP.A ELISA method to detect HIV-1 p24 antigen was established.Eighty eight HIV positive sera and 50 normal sera were detected simultaneously by Dopont HIV-1 p24 antibody reagent and the ELISA method established by our laboratory to confirm its specificity.Results The results of detecting HIV-1 p24 antigen of 88 positive sera and 50 normal sera by two reagents were all negative,which showed that the coincident rate was 100%.The sensitivity of our reagent to detect HIV-1 p24 antigen was 100pg/ml.Conclusion The ELISA methods of detecting HIV-1 p24 antigen has been preliminary established.Our reagent has good sensitivity and specificity as compared with imported ones,which has laid an important basis for developing a more sensitive diagnostic reagent kit for detecting HIV-1 p24 antigen.