The aim of this study was to evaluate the heat-dissociated p24 antigen (HD p24 Ag) assay as an alternative low-cost tool for diagnosis of HIV-1 infection and quantitation of HIV-1 RNA levels in African adults mainly infected with HIV-1 CRF02_AG strains. One hundred seventeen plasma specimens were obtained from HIV-1-seropositive subjects enrolled in the ANRS 1220 PRIMO-CI cohort (Abidjan, Côte d'Ivoire, West Africa). Results of the HD p24 Ag assay were comparable with those of HIV-1 RNA levels quantified in the same antibody-positive plasma samples by the Amplicor HIV-1 Monitor assay (1.5 version; Roche Diagnostics, Indianapolis, IN): sensitivity, 95.7% versus 96.6%, respectively; specificity (evaluated with samples from 75 seronegative subjects), 94.7% versus 100%, respectively. HD p24 Ag and HIV-1 RNA assays were weakly correlated (Spearman coefficient correlation, r = 0.33; P < 0.001) except for HIV-1 RNA levels of >/=5 log10 copies/mL (r = 0.62; P < 0.001). Quantitation of HD p24 antigenemia in 76 plasma specimens from 14 patients treated with highly active antiretroviral therapy demonstrated weaker changes during treatment than those observed with the HIV-1 RNA assay. Follow-up of infected patients using both markers showed different results. The reliability of the HD p24 Ag assay is questionable for clinical and biologic management as a surrogate tool for measurement of HIV-1 RNA levels in Africa.