Background: The current majority of modern medicine is to prevent infection due to hospital acquired microbes and evolving bacterial organisms which can affect the nursing staff due to improper hygiene. In this study we sought to study the practice of hand washing among nursing staff in tertiary care teaching hospital. Material & Methods: A cross–sectional study was conducted to assess hand washing practice among the nursing staff in tertiary care teaching hospital. Results: A total of 150 nursing staff were studied for their practice of hand washing. It was found that a total of 77% staff practiced hand washing before examining patient and a total of 90% staff practiced handwashing after examining the patients. Compliance with hand washing practice was observed more in B.Sc and M.Sc qualified nurses than in ANM and GNM qualified nurses. Conclusions: The majority of nursing staff wash their hands after the patient care. Educational qualification and Service experience of healthcare professionals are showing impact on the hand hygiene practice. The current study emphasizes the need of counselling on hand hygiene to the healthcare professionals. Hence it is important to conduct regular training programs on hand hygiene for nurses with continuous monitoring.