Background. To investigate any epidemiological association between human herpesvirus (HHV)-8 and prostate cancer, we determined the prevalence of HHV-8 seropositivity among prostate cancer case and control subjects in the United States and Trinidad and Tobago.Methods. Antibodies against HHV-8 were detected in 2 independent laboratories using either indirect immunofluorescence assay (IFA) or a combination of enzyme-linked immunosorbent assay and IFA.Results. Among 138 Tobago men with prostate cancer, HHV-8 seroprevalence was 39.9%-significantly higher than that among 140 age-matched control subjects (22.9%; P=.003; odds ratio [OR], 2.24; 95% confidence interval [CI], 1.29-3.90). Among 100 US men with prostate cancer, seroprevalence was 20%-significantly higher than that of 177 blood donors (5.1%; P=.001; OR, 4.67; 95% CI, 1.91-11.65) and higher than that of 99 men with cancer not related to HHV-8 (13%; P=.253; 95% CI, 0.77-3.54).Conclusions. HHV-8 seropositivity is elevated among men with prostate cancer compared with control subjects, which suggests that HHV-8 plays a role in the development of prostate cancer.