ABSTRACT Surgical aseptic technique is the application of those strict principles and procedures aimed at preventing the invasion of surgical sites by micro organisms. Prevention of surgical site infection (SSI) has become a priority worldwide. Approximately 50,000 deaths are estimated to occur annually across Europe, as a consequence of surgical site infection. In some university teaching hospitals in Nigeria, the rate of SSI ranges from 10 to 20%. This was linked to non compliance with standard practice due to poor theatre workers’ attitude to work. Compliance to surgical aseptic standard in this study entails theatre workers’ readiness and obedience to carry out the correct procedure in the area of surgical hand scrub, proper use of personal protective device and maintenance of sterile field during surgery. The purpose of this study is to determine the attitude and compliance of theatre workers towards the practice of surgical aseptic standard in tertiary health institutions in Enugu state. The specific objectives were to: (i) elicit theatre workers’ attitude towards various aspects of surgical aseptic standard, (ii) determine theatre workers’ practice of surgical aseptic standard, (iii) determine the relation between attitude and compliance in the practice of surgical aseptic standard and (iv)compare the practice of standard aseptic technique across tertiary health institution in Enugu State. The study adopted a descriptive cross sectional survey design. It was conducted at the three tertiary health institutions in Enugu State, Nigeria. Using purposive sampling technique and observing the following inclusion criteria: (a) participants must be doctors and nurses working in the operating theatre, (b) must be certified, registered, and licensed to practice in the operating theatre, (c) must be physically fit to operate or assist during surgery and (d) willingness to participate in the study. A total of 272 theatre workers were recruited from the population of 660 theatre worker. Instruments for data collection were the pre tested, 20 item researcher developed questionnaire on attitude of theatre workers and 40 item observational checklist. The instruments were validated by the researcher’s supervisor and two experts in the Department of Nursing in University of Nigeria Enugu Campus. Reliability of the questionnaire was established using test retest method at two weeks interval that yielded Cronbach’s alpha correlation coefficient of 0.88. The observational checklist, inter rater scores showed high degree of agreement. Data collected were subjected to descriptive statistics and analysed using Pearson’s Product Moment Correlation and analysis of variance (ANOVA) at p < 0.05 significant level. The significant means decision point for attitudinal item is >2.5 and >2 for observational checklist. The perceived attitude of theatre workers to surgical aseptic technique was positive with overall mean of 3.40 ± 0.81, while SHS had 3.79 ± 0.18, use of PPD 3.1 ± 0.18, and MSF had 3.43 ± 0.81.Theatre workers’ compliance to surgical standard practices were as follows: surgeons and peri operative nurses consistently had higher mean score, for SHS (2.89 ± 0.19, 2.78 ± 0.18), use of PPD (2.84 ± 0.12, 2.81 ± 0.22) and MSF(2.75 ± 0.20, 2.84 ± 0.19) respectively, than physician and nurse anesthetists score SHS (2.34 ± 0.45, 2.39 ± 0.40), use of PPD (2.43 ± 0.35, 2.71 ± 0.31) and MSF (2.29 ± 0.42, 2.61 ± 0.41) respectively. There were significant relationship (r= 0.86, r= 0.81, r= 083) between attitude and compliance. There were no significant variations (p> 0.05) in the practices across the three health institutions. Surgeon and physician anesthetists showed less compliance to the practice of wearing theatre outfit within the restricted area, with mean score of 1.67 and 1.56 respectively.