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Background and Objectives: Patient safety in hospital is an essential professional indicator that should be noticed. The thread of fires is of the most potentially dangerous risk that could harm patients and personnel. Hence, we studied operating room staff knowledge about fires, its prevention, and control, based on their specialty and their job tenure. Materials and Methods: This is a descriptive-analytical cross-sectional study. A cluster sampling method was applied, and a quota was considered for each hospital. In each cluster, samples were selected through census method. Data were collected using a three-part questionnaire and analyzed using SPSS version 17 (SPSS, Inc, Chicago, IL, USA). Results: The mean of their knowledge level about fires and its prevention was 2.71 (0.67), and control was 2.62 (0.71). In 66% of cases, the level of knowledge about fires and its prevention, and in 70% of cases, the level of knowledge about methods of fire control have been below the average (P < 0.05). As staff age increases, their level of knowledge increases as well. Hospital they are employed by, their job tenure, and their profession can affect the level of knowledge (P < 0.05). Discussion: This study shows that the staff knowledge about fires, its prevention, and control has been lower than the average limit. Due to the sensitivity of the issue, in terms of susceptibility of the environment and medical equipment for ignition, and also the safety of staff and patients, it is necessary to make facilities for development of staff knowledge, to eliminate defects of staff training by managers, and also to consider standards.