Detection of early anti-p24 HIV responses in EIA- and immunoblot-negative individuals. Implications for confirmatory...

A sensitive and specific radioimmunoprecipitation assay was developed for the detection and analysis of anti-HIV antibody response in human sera with the use of 125I-labelled purified HIV proteins with subsequent sodium-dodecylsulfate gel electrophoresis (125I-RIPA). The 125I-RIPA was shown to be as specific but at least 1 log more sensitive with respect to the detection of gp41env and p24gag than the immunoblot analysis as tested in serum samples from several risk groups. Sequential sera were obtained from 9 individuals who seroconverted for HIV antibodies. In 4 individuals, antibody to p24gag was detected in earlier serum samples by the 125I-RIPA than by EIA or immunoblot; in the other 5 individuals, the detection of p24gag concorded in enzyme-linked immunosorbent assay (EIA), immunoblot and 125I-RIPA. Moreover, in one of 78 randomly chosen EIA-negative sera from individuals at high risk, antibodies to p24gag could be detected by the 125I-RIPA. This early seroconversion was confirmed 3 months later by means of immunoblotting and EIA. The specificity of the 125I-RIPA was further demonstrated by analyzing sequential EIA-negative serum samples from 10 individuals at risk for AIDS, collected during 2 years at 3-monthly intervals. All 80 serum samples were found to be negative in the 125I-RIPA and the individuals revealed no signs of HIV infection. The 125I-RIPA technique may be a valuable confirmatory assay in the serology of HIV infections. The sensitivity of this test provides a reliable measure of effective sensitivity when new-generation screening tests are evaluated.(ABSTRACT TRUNCATED AT 250 WORDS)