Introduction: Health care workers (HCW) may avoid healthcare-associated infections through adherence to Standard Precautions (SP) resulting in better clinical outcomes and decreased medical costs. Reduction of that must be considered for all blood borne pathogens through adherence to standard precautions, using personal protective equipments, appropriate use of safety devices, and providing a needle disposal system in the work place. Some articles have involved data about adherence of healthcare workers to standard precautions toward infection control and prevention in developing countries. Aim of work: To assess Knowledge, Attitude, and Practices of HCW to SP of infection control guidelines in one of the major tertiary health care facilities in the Eastern Province, Saudi Arabia. Materials and methods: A cross-sectional study was conducted to asses Knowledge, Attitude, and Practices related to SP among 202 medical staff; physicians and nurses, using a standardized self-administered questionnaire based on CDC (Center for Disease and Control) guidelines to Standard Precautions from May to July 2015. Results: physicians achieved higher score of Knowledge compared to nurses (P<0.05). The mean score of Knowledge, Attitude and Practices among females is higher than males with statistically significant difference, and higher scores of Knowledge were associated with higher scores of Practices. Conclusion: Findings of the present study highlighted the need for continuing training programs to target nurses and physicians to establish acceptance of appropriate practices that enable them to adopt and adhere to SP.