INTRODUCTION: Hand hygiene plays a key role in the prevention of healthcare-related infections. Healthcare associated infections lead to prolonged hospitalization, additional treatment, increased healthcare costs, disability and mortality. Contaminated hands of healthcare workers are the most common cause of healthcare associated infections. OBJECTIVE: The aim of this thesis was to examine the knowledge level of the students of the third, fourth and fifth grades of the Ante Kuzmanić Zadar High School of Nursing and compare the obtained results with the results of foreign research. The aim of the study was to examine whether there are differences in the knowledge of hand hygiene among third-grade students compared to fourth and fifth grade students of the Ante Kuzmanić Zadar Nursing School. RESPONDENTS AND METHODS: The subjects were students of the third, fourth and fifth grades in the Ante Kuzmanić High School of Nursing, Zadar. The total number of students who participated in the research was 125 (N=125). A standardized questionnaire on hand hygiene knowledge for medical staff was used in dana collection, which was downloaded from the World Health Organization website. RESULTS: 33 students attended third, 44 fourth and 48 fifth grade students. Dana analysis indicates that 80,8% of students have an average level of knowledge, while 8% of students have a poor level of knowledge about hand hygiene. Also, through the research, was found that there no difference in the level of knowledge about hand hygiene among third graders compared to fourth and fifth graders. CONCLUSION: Students of the third, fourth and fifth grades have middle level of knowledge about hand hygiene, which results ware consistent with the results of available foreign research. There was no statistically singnificant difference in the level of knowledge between third grade students and fourth and fifth grade students. It was found that third, fourth and fifth grade students have the same level of knowledge about hand hygiene.