Abstract Background: For better planning, hospital infection control supervisors should be aware of the behavioral status of their employees. The aim of this study was to evaluate the knowledge, attitude and practice of nurses and physicians in the field of infection control at a tertiary university hospital in north-eastern Iran. Methods: In this prospective cohort study, a researcher based questionnaire was used to assess the knowledge and attitude of the participants. Additionally, a checklist completed by several trained observers, was used to evaluate their practice, based on the opportunities and overall performance of the subjects. Results: Totally 334 people (including 180 nurses and 154 physicians) were evaluated. Only 1.5% of respondents had low knowledge level. There was no significant difference between the knowledge level of physicians and nurses (p= 0.101). Also, 97.7% of subjects had a positive attitude towards infection control issues. In general, poor practice was only detected in 8.7% of the participants. Nevertheless, in issues such as hand hygiene upon entering the ward, removing rings or watches during hand washing and hand washing with soap and water for 15-30 seconds, less than 20% of them had proper performance. Conclusion: The present study demonstrated that the knowledge and attitude of physicians and nurses towards infection control issues in hospital seem acceptable. However, the percentage of poor practice compared with attitude and knowledge was higher. Despite overall favorable practice, the participants’ performance was poor with respect to certain important issues that deserve special attention. Keywords: Nosocomial infection; Infection control; Knowledge; Attitude; Practice

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