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Epidemiologic studies suggest that human herpesvirus 8 (HHV-8) may be sexually transmitted. To study the potential for HHV-8 transmission through cervicovaginal (CV) secretions, the presence of HHV-8 DNA was investigated by nested polymerase chain reaction in the cellular fraction of CV secretions from 36 human immunodeficiency virus type 1 (HIV-1)-seropositive and 29 HIV-1—seronegative women. The same patients were tested for antibodies to two defined HHV-8 antigens (latency-associated nuclear antigen and open-reading frame 65-encoded structural protein) and for HHV-8 DNA in their peripheral blood mononuclear cells (PBMC). The findings were compared with the rate of HHV-8 detection in semen samples of 20 HIV-1—infected men. HHV-8 DNA was detected in the CV samples from only 1 HHV-8-seropositive AIDS patient, in 3 PBMC samples (1/29 HIV-1—seronegative patients, 1/3 AIDS patients with Kaposi's sarcoma, and 1/19 AIDS patients), and in 1 of 20 semen samples. HHV-8 infection was more common in HIV-1—infected than uninfected women. Thus HHV-8 DNA is only rarely detectable in CV secretions and semen of HHV-8—infected individuals.