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Prostitution may constitute a risk behavior for infection with hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus (HIV). We conducted a seroepidemiologic study among 200 male transvestite commercial sex workers (CSWs) in Montevideo, Uruguay in 1999. Evidence of exposure to HBV, HCV, and HIV was found in 101 (50.5%), 13 (6.5%), and 43 (21.5%) individuals, respectively. Positivity for HIV was correlated with both HBV (odds ratio [OR] = 2.15, 95% confidence interval [CI] = 1.01–4.67) and HCV (OR = 3.47, 95% CI = 0.90–12.79) infection. Predictors of infection were older age (≥ 26 years; P < 0.01) for all 3 viruses and time in CSW (≥5 years; P < 0.05) for HBV and HIV. Prior history of use of drugs (OR = 3.54, 95% CI = 1.09–11.52) and sexual contact with foreigners (OR = 9.2, 95% CI = 1.16–73.12) were found to be associated only with HCV infection. Sexual transmission of these viruses constitutes a significant problem among male transvestite CSWs.