Needlestick injury is one of the significant stress-leading causes among nurses in healthcare settings. Aim: The aim of this study is to assess the level of stress caused by fear of needlestick injury exposure among nurses working at Sohag University Hospital. Subject and methods: A cross-sectional study conducted with on 178 nurses from different Sohag University Hospital departments using two tools, demographic and needlestick information tool and stress level tool. Results: About seventy percent of nurses from a variety of units in Sohag University Hospital suffered from severe stress of exposure to needlestick injuries and indicated that they have been actually exposed to prickle in the preceding two months. Majority of injuries occurred in the morning shift at ICU, surgical, medical and their belonging subspecialties departments by syringes needles, IV devices and medical scalpels. The respondents graduated from Secondary School of Nursing were the most common were highly exposed to needlestick injuries, and were planning to leave the clinical nursing procedures or their changed career categorically. A significant percent of nurses preferred not to report official authorities on needlestick injuries for these reasons, busy job, no possibility of infection from needlestick, and no way to communicate with officials. The most common procedures for needlestick injuries occur among nurses was during recapping of needles, preparing and giving drugs and most of them did not perform any serological tests after injuries occurred. In addition, the current study shows that the high percent of nurses washed the injured place by the running water and soap, they used the disinfectants material to clean the injury site and put a dressing on the wound, and the most common sites of injury were the hands and fingers, and it occurred once or twice. There was a statistical significant correlation among needlestick injuries and educational level, departments, leaving of specific clinical nursing procedures and level of stress while no statistically significant relation is found between the previous training and needlestick. Conclusion & Recommendations: Exposure to needlestick injuries may cause severe stress among the nursing staff. Appropriate training and awareness should be given regularly to the nurses to ameliorate skills to deal with needlestick injuries during their nursing curriculum and continue after graduation to help reduce stress among nurses.