Background and aims: The operating room is a critical place for accident. One of the events that is likely to occur at this location is fire accident during the surgery. This type of accident rarely happens, but if happen, they can lead to serious injury or death of patients. Therefore the purpose of this study is the assessment of awareness and practice of operating room personnel about prevention of fire and effective factors in operating room in Khuzestan hospitals. Methods: This cross-sectional descriptive study was conducted among operating rooms in Khuzestan hospitals in 2014. The sample consisted of 253 employees (31% anesthesia nurse and 69% operating room nurse) from 18 hospitals who were selected by simple random sampling. In this study, all operating rooms personnel were investigated, in random sampling. Fire prevention questionnaires were used to collect information; awareness and practice of operating room personnel about preventing of fire, the relationship between these two factors with the age, work experience and the effect of training, education, sex and job title were also investigated. To examine the relationship between variants, SPSS v. 16 was used and for comparing the groups Mann– Whitney was used. Results: Considering awareness of strategies of preventing fire, 15% had high awareness, 25% medium, 48. 4% poor and 11. 1% had no awareness. For operation, 87. 2% were high operating, 14. 2% medium, 6. 7% poor and 8% were very poor. There was no relation between the age and work experience with awareness and operation in Kendal's correlation test (p>0. 05). The comparison test in this study showed that training is effective on operation and awareness (p<0. 05), but there was no significant difference between awareness and operation of man and women. Also, there was a significant difference in comparison of average scores of awareness between education levels (technician and bachelor) (p<0. 05), but there was no significant differences between the type of jobs (p>0. 05). This study showed that the level of operation of operating room personnel was more than anesthesia personnel and the differences was significant (p<0. 05). Conclusion: The results from this study and previous ones indicate that awareness about fire prevention ways between operating room personnel is low and their safety is not satisfied. However, performance is at the appropriate level to fire prevention. The result of this study also showed that factors such as previous training in the field of fire prevention, education level and type of job can be factors related to awareness and practice toward prevention of fire.