Purpose: Hospital-acquired infections, common in developing countries, are a leading cause of morbidity and mortality. This study was done to examine the effects of organizational culture and awareness of infection control on nursing practice of infection control by nurses working in emergency departments. Methods: This descriptive correlational study was conducted with 135 nurses from the emergency department of a hospital. Data were collected from December 23, 2019, to January 14, 2020, using a structured questionnaire. Collected data were analyzed using descriptive statistics, t-test, ANOVA, Pearson correlation coefficients, and multiple regression analysis with SPSS/WIN 18.0. Results: The mean score of organizational culture, awareness, nursing practice of infection control were 5.56±0.78 out of 7, 4.66±0.36 out of 5 and 4.51±0.38 out of 5, respectively. The multiple regression analysis demonstrated that awareness and organizational culture of infection control accounted for 43% of the nursing practice of infection control. Conclusion: Study findings showed that there is a significant correlation between organizational culture, awareness, and nursing practice of infection control in the emergency department. Therefore, to improve nursing practices infection control in the emergency department, efforts should be made to promote infection control by developing education programs that can improve infection control awareness as well as strategies for a positive organizational culture.