Saliva and blood samples were tested for human immunodeficiency virus-1 (HIV-1) antibodies in two high-risk populations in Kinshasa, Zaire. In a seroprevalence study of 458 sexually transmitted disease (STD) clinic attendees, 142 of 145 seropositive individuals had enzyme-linked immunosorbent assay (ELISA)-positive saliva samples (97.9% sensitivity). All saliva samples from seronegative patients were ELISA-negative (100% specificity). Of the 142 ELISA-positive saliva specimens, 137 were also Western blot-positive (94.5% sensitivity). In a subsequent seroincidence study of 315 initially seronegative female prostitutes followed during 183 woman-years of observation, 9 of 14 women who seroconverted (7.7% seroincidence) had ELISA-positive saliva samples at the time seroconversion was detected. Only three of these saliva specimens could be confirmed by Western blot. Although salivary testing for HIV-1 antibodies using conventional assays was not sensitive in detecting recent seroconversions, screening of salivary samples for HIV-1 antibody provides a convenient alternative method for conducting seroprevalence surveys in populations in whom venipuncture is not possible or convenient.