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Victims could become infected with sexually transmitted infections (STIs) during a sexual assault. Several guidelines recommend presumptive antimicrobial therapy for sexual assault victims (SAVs). We assessed the STI positivity rate and treatment uptake of female and male SAVs at the Amsterdam STI clinic.Sexual assault victims answered assault-related questions and were tested for bacterial STI (chlamydia, gonorrhea, and syphilis), hepatitis B, and HIV during their initial visits. Sexual assault victim characteristics were compared with non-SAV clients. Backward multivariable logistic regression analysis was conducted to assess whether being an SAV was associated with a bacterial STI. The proportion of those returning for treatment was calculated.From January 2005 to September 2016, 1066 (0.6%) of 168,915 and 135 (0.07%) of 196,184 consultations involved female and male SAVs, respectively. Among female SAVs, the STI positivity rate was 11.2% versus 11.6% among non-SAVs (P = 0.65). Among male SAVs, the STI positivity rate was 12.6% versus 17.7% among non-SAVs (P = 0.12). In multivariable analysis, female SAVs did not have increased odds for an STI (odds ratio 0.94; 95% confidence interval, 0.77–1.13), and male SAVs had significantly lower odds for an STI (odds ratio, 0.60; 95% confidence interval, 0.36–0.98). Of SAVs requiring treatment, 89.0% (female) and 92.0% (male) returned.The STI positivity rate among female SAVs was comparable with female non-SAVs, but male SAVs had lower odds for having a bacterial STI than did male non-SAVs, when adjusting for confounders. The return rate of SAV for treatment was high and therefore does not support the recommendations for presumptive therapy.