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Objective To describe the characteristics of female sex workers (FSW) in 12 Brazilian cities. Methods This was a cross-sectional biological and behavioral surveillance study with FSW recruited by respondent-driven sampling (RDS) in 2016. HIV prevalence, sociodemographic, behavioral and sex work profession variables were estimated by city. The chi-square test was used to compare these distributions. Results Among 4,328 FSW, HIV prevalence was 5.3% (95%CI 4.5;6.3), ranging from 0.2% (95% CI 0.0;1.2), in Campo Grande, to 18.2 (95%CI 13.2;