Driving after a night shift imposes a risk on health care professionals and other road users. The aim of this study was to measure psychomotor performance of driving of night shift nurses compared to day-shift nurses.Forty-seven volunteer female nurses working at Sina hospital in Tehran, Iran, with a call in all departments of hospital, participated in this study (23 night shift and 24 day shift nurses) in 2016. The tests included RT for simple reaction time, ATAVT for perceptual speed, LVT for visual orientation and ZBA for time anticipation. Data collection tools were individual characteristics, 11-item circadian type inventory (CTI), Stanford sleepiness scale (SSS), and Swedish occupational fatigue inventory (SOFI-20) questionnaires. Psychomotor driving performance was assessed using validated computerized traffic psychological battery of Vienna Test System (VTS), before and after the shifts. Data analysis was performed using paired-samples t-test and Linear Regression.The mean age of day and night-shift nurses were 31.4±5.6 and 28.7±3.9 years respectively, no significant difference between two groups. Thirty percent of night shift and 16.7% of day shift nurses reported traffic accidents in the past year. The results revealed that, scores based on viewing times in visual orientation test (p=0.005), and median reaction time score in choice reaction time and reactive stress tolerance test (p=0.045), had a significant association with a 12-hour night shift with a 3-hour nap.Twelve-hour night shift work impairs choice reaction time and visual orientation in nurses, even though they take a 3- hour nap during the shift. These skills are required for safe driving.