Background & Aims: Health care-associated infections (HCAIs) are a major patient safety concern, and consequently prevention should be a top priority for institutions. Hand hygiene is the most effective way to reduce nosocomial infections. Materials & Methods: 296 medical students in different wards of university hospitals in Urmia participated in this descriptive study. They completed a hand wash checklist and a hand hygiene questionnaire. Finally, the obtained data were analyzed using SPSS25 software. Results: The results showed that 48% were male and 52% were female with an average age of 24.5 ± 2.9 years. Most of the participants were recruited from Imam Khomeini Hospital (%35) while only%5/8 of the participants were recruited from Razi Hospital. 34.7% of the participants were extern, 44.6% were intern and 20.6% were resident. The average level of awareness of hand hygiene was 47%, the minimum level of awareness was 31% and the maximum level of awareness was 69%. The average attitude toward hand hygiene principles among medical students was 31% (weak), the lowest attitude was 22% (weak) and the highest attitude was 58% (good). The average performance of hand hygiene practice principles was 39% (weak), the lowest was 22% (weak) and the highest was 51% (good). The average level of awareness of hand hygiene principles among female and male medical students were 50% and 44%, respectively. The average attitude among female medical students was 40% and average attitude among male medical students was 26%. The average performance of the principles of manual health practice among female and male medical students were 26% and 23% (acceptable), respectively. The average level of awareness, attitude, and performance of hand hygiene principles among students of external course, internship, and residency were 51%, 40% and 35%; 45%, 28% and 17%,; and 47%, 38% and 25%, respectively. In general, the average performance of hand hygiene principles among students in the ICU, surgical department and internal medicine department were 48%, 40%, and 27%, respectively. Conclusion: Due to low-level attitude and knowledge of medical students regarding hand hygiene and the necessity of hand hygiene in preventing infection, as well as the importance of patient health during the hospitalization period, it is essential to hold in-service training classes in the field of hand hygiene to improve and promote hand hygiene.