Introduction: Sexually transmitted infections (STIs) incidence, as most common acute diseases in the world, has been increasing. We investigated knowledge, attitude, and practice (KAP) of healthcare staff, including physicians and healthcare providers, such as health professionals and midwifes. Material and methods:In this cross sectional study, self-made questionnaire was applied. Data collection was performed between September and November 2019. Participants were selected using stratified random sampling method, and 226 healthcare staff were included in the study. Mann-Whitney and Kruskal-Wallis tests were used to compare median and interquartile range (IQR) score of KAP among participants. KAPs were classified into poor, moderate, and desirable. Data were analyzed in SPSS version 21.Results: Total median (interquartile range) score of KAP was 14 (4), 29 (8), 30 (9), respectively. Knowledge score of health professionals were low (52%), midwives and physicians scored average (74% and 75%, respectively). Most health professionals, midwives, and physicians scored moderate on attitudes (61%, 78%, and 80%, respectively). The results indicated that 60% of midwives performed well, with 46% of health professionals and 45% of physicians having mediocre performance. Conclusions:In this study, health professionals performed well, but their level of knowledge was low. This means that they perform well according to their duties, but did not have enough knowledge. Therefore, they need more training to improve the quality of their services.