Background: Gowning and gloving is regarded as the basic component of modern Surgery, its standard of practice is directly concerned with patient safety and infection prevention. The aim of this study was to observe the practice of gowning and gloving technique among health personnel at operation theatre of teaching hospital, Bharatpur, Nepal. Methods: A descriptive cross-sectional study was carried out among 90 health personnel who had scrubbed in the operation theatre before approached to sterile field, by using observational checklist. Sample was selected by using non probability, consecutive sampling technique. Data were analyzed using descriptive and inferential statistics to find the association between the variable. Results: The findings highlighted, that only 36.7% of the health personnel fulfill the criteria of surgical gowning and gloving. The main issues found in the study were not picking up the entire folded gown through all layers from the wrapper (31.1%), and not holding the gown near the gown’s neck and touch unsterile objects (18.9%). Conclusions: The level of practice on gowning and gloving technique was statistically significant with professional experience and education of the respondents. Hence, operation theatre department should develop the audit protocol and in-service training model for the surgical team.