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Objective: To evaluate the prevalence of hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus (HIV) among pregnant women in Puerto Rico., Methods: An anonymous serosurvey was conducted in four prenatal clinics in San Juan, Puerto Rico, involving women presenting consecutively for their first prenatal visit., Results: Nineteen of 997 pregnant women (1.9%, 95% confidence interval [CI] 1.2-3.0) tested positive for HCV antibody (anti-HCV), and eight (0.8%, 95% CI 0.4-1.6) were HIV seropositive. Of the 992 women for whom serum samples were tested for HBV markers, 91 (9.2%, 95% CI 7.5-11.2) had evidence of past or current HBV infection, and four (0.4%, 95% CI 0.1-1.1) were HBV carriers. The age-specific HBV prevalence ranged from 4.1% among women 15-19 years old to 18.5% among those at least 30 years old (P < .001, chi 2 test for trend). Anti-HCV prevalence was also higher among women at least 30 years old compared to younger women (3.1 versus 1.9%; prevalence ratio 1.6, 95% CI 0.6-4.9), although the difference was not statistically significant. Anti-HCV prevalence was higher among women with past or current HBV infection than among women who were not infected (7.7 versus 1.3%; prevalence ratio 5.8, 95% CI 2.3-14.3)., Conclusions: The prevalence of chronic HBV and HCV infection among pregnant women tested in San Juan, Puerto Rico, is comparable to that among pregnant women in the United States. The prevalence of HIV infection among pregnant women in San Juan is higher than among childbearing women in the United States.