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To assess the role of sexually transmitted infection (STI) management to prevent HIV acquisition among sex workers in Burkina Faso.Open cohort study of professional and nonprofessional sex workers with 3-month follow-up visits.Baseline and follow-up visits consisted of the administration of a behavioral questionnaire, education sessions on HIV and STIs, a medical examination, and laboratory testing for STI and HIV diagnosis.Three hundred seventy-seven HIV-negative women were enrolled in the study. The cumulated HIV incidence was 3.2 per 100 person-years (Poisson 95% confidence interval: 1.9-4.9). Bacterial and parasitic STIs were low at baseline, whereas herpes simplex virus-2 (HSV-2) prevalence was 54.7%. By a Cox regression model, self-assessment of high HIV risk in the past, less than 5 clients per week, and no change of a steady partner were independently associated with HIV acquisition. Among STIs, only infection with HSV-2 tended to be associated with HIV acquisition (odds ratio = 2.45; P = 0.15). Overall, condom use increased during the study but to a lesser extent with steady partners.Bacterial and parasitic STIs are no longer a key determinant of HIV acquisition, given the current stage of the outbreak in Burkina Faso. Although efforts for STI control should be maintained, strategies should focus on nonprofessional sex workers, steady partners, and HSV-2 infection to tackle HIV transmission further in this high-risk group.