Background: Surgical asepsis practices should maintained by the surgical team to prevent contamination of the open surgical wound. Aim: This study aimed to evaluate the surgical asepsis practices in the operating theatre of King Khalid Hospital, Najran. Methods: The researchers utilized a three data collection forms to collect the needed data about practices of surgical asepsis. Swabs obtained from surgical site and from OR surfaces for bacteriological examination. Results: only 55% of patients showered the day of surgery, there was no cleaning of light and scrubbing sinks at 60% and 75% of surgeries respectively. Sterile persons touch only sterile items at 55% of surgeries, 84.9% of them performed surgical scrubbing correctly. There was a contamination of OR floor and conditioning system in 55% of surgeries, there was a surgical site infection(SSI) after suturing among 40% of patients and there was a correlation between contamination of scrubbing taps and SSI post- preparation (P < 0.05). Conclusion: There was an improper surgical asepsis practices performed by OR surgical team, we recommend the importance of continuing education among OR staff to keep them updated with the new trends and developments in surgical asepsis.