Objectives: This study was conducted to assess hospital staff’ knowledge, selfreported compliance and actual practice of universal precautions, identify important non adherence factors, and determine the proportion of participants exposed to blood and other body fluids with identification of the associated risk factors. Subjects and Methods: A total of 152 hospital staff from different job categories at risky departments in a tertiary care hospital in Ismailia City were included in this survey study where a structured questionnaire and a direct observation check list were used. Results: The mean percent score of participants’ self-reported compliance was much higher than those of knowledge and actual practice. Moreover, a statistically significant positive correlation was found only between percent score of participants’ knowledge and practice. A gab between knowledge and practice was detected where 75% of the studied participants had insufficient knowledge while 36.2% of them had insufficient practice.Insufficient knowledge was significantly revealed among males, nurses and surgical departments’ staff; while insufficient practice was significantly detected among nurses, surgical departments’ staff, those with > 48 working hours per week and hospital staff with > 3 constrains. The majority of the participants (65.1%) got their information from sources other than the hospital. Work-related factors were the most important compliance barriers. The majority of the participants had history of sharps injury and exposure to blood or other body fluids splashes at work; however, none of these Objectives: This study was conducted to assess hospital staff’ knowledge, selfreported compliance and actual practice of universal precautions, identify important non adherence factors, and determine the proportion of participants exposed to blood and other body fluids with identification of the associated risk factors. Subjects and Methods: A total of 152 hospital staff from different job categories at risky departments in a tertiary care hospital in Ismailia City were included in this survey study where a structured questionnaire and a direct observation check list were used. Results: The mean percent score of participants’ self-reported compliance was much higher than those of knowledge and actual practice. Moreover, a statistically significant positive correlation was found only between percent score of participants’ knowledge and practice. A gab between knowledge and practice was detected where 75% of the studied participants had insufficient knowledge while 36.2% of them had insufficient practice. Insufficient knowledge was significantly revealed among males, nurses and surgical departments’ staff; while insufficient practice was significantly detected among nurses, surgical departments’ staff, those with > 48 working hours per week and hospital staff with > 3 constrains. The majority of the participants (65.1%) got their information from sources other than the hospital. Work-related factors were the most important compliance barriers. The majority of the participants had history of sharps injury and exposure to blood or other body fluids splashes at work; however, none of these Objectives: This study was conducted to assess hospital staff’ knowledge, selfreported compliance and actual practice of universal precautions, identify important non adherence factors, and determine the proportion of participants exposed to blood and other body fluids with identification of the associated risk factors. Subjects and Methods: A total of 152 hospital staff from different job categories at risky departments in a tertiary care hospital in Ismailia City were included in this survey study where a structured questionnaire and a direct observation check list were used. Results: The mean percent score of participants’ self-reported compliance was much higher thanthose of knowledge and actual practice. Moreover, a statistically significant positive correlation was found only between percent score of participants’ knowledge and practice. A gab between knowledge and practice was detected where 75% of the studied participants had insufficient knowledge while 36.2% of them had insufficient practice. Insufficient knowledge was significantly revealed among males, nurses and surgical departments’ staff; while insufficient practice was significantly detected among nurses, surgical departments’ staff, those with > 48 working hours per week and hospital staff with > 3 constrains. The majority of the participants (65.1%) got their information from sources other than the hospital. Work-related factors were the most important compliance barriers. The majority of the participants had history of sharps injury and exposure to blood or other body fluids splashes at work; however, none of these incidents were reported. These exposures were significantly revealed among nurses, staff in surgical departments and those with insufficient knowledge and practice of universal precautions. Conclusions: Hospital staff are at high risk of exposure to blood borne pathogens, yet universal precautions are not well understood or implemented. So, implementing a specifically tailored infection-control program will be most effective in protecting them. .