Live-cell indirect immunofluorescence was used to detect HIV-associated antigens in fresh uncultured peripheral blood mononuclear cells from 29 HIV-seropositive subjects. Conventional fixed-cell immunofluorescence and live-cell immunofluorescence were run in parallel on each sample using monoclonal antibodies directed against HIV gag and env gene products. Sera from the same patients were also tested for HIV antigen by a sandwich enzyme immunoassay. Results strongly indicate that nondenaturing live-cell immunofluorescence is much more sensitive for detection of HIV-associated antigens and may be useful in the diagnosis of HIV infection as well as in the study of pathogenesis.