Introduction: Knowing the factors associated with periodic HIV testing among female sex workers (FSW) is essential to expand testing coverage and to broaden programs of treatment as prevention. Methods: We used data from 4,328 FSWs recruited by the respondent-driven sampling (RDS) method in 12 Brazilian cities in 2016. Data analysis considered the complex sampling design. The prevalence of HIV testing in the last year and periodic HIV test were estimated. Factors associated with regular HIV testing were identified through logistic regression models. Results: The testing coverage in the last year was 39.3%. Only 13.5% of FSW reported having performed a periodic HIV test in the last year. Among the factors associated with the higher probability of HIV testing in the last year were a better level of education, living with a partner, working indoors, consistent use of condoms, and regular use of public and private health services stood out. Discussion: Periodic HIV testing allows early diagnosis and immediate treatment of cases, reducing the chances of spreading the infection to the population. However, factors such as stigma and discrimination hinder the use of regular health services. Conclusion: It is necessary to expand awareness campaigns, especially among FSWs with low educational level and greater vulnerability, in order to broaden the perception of risk and the importance of periodic testing, in addition to encouraging regular health care.