Objectives: The objectives of this study were to determine and compare the knowledge, attitudes and practice of preventing the occurrence of nosocomial infections among students of the Faculty of Medicine in Osijek, and to determine whether there is a difference regarding age, sex and study program. Study design: Cross-sectional. Participants and Methods: The study involved students from three different study programs at the Faculty of Medicine in Osijek (Medicine, Nursing, Medical laboratory diagnostics) that have completed a course with the subject of nosocomial infections as part of the curriculum during the academic year 2015/2016 (Medical Microbiology and/or a course about infectious diseases). Results: The study included data from 316 respondents. It was found that the knowledge of students of the Faculty of Medicine in Osijek kept up with the global trends of similar surveys conducted in other European countries. The difference in relation to age existed among the group of students from 22 to 24 years of age, regarding the knowledge about standard precautions. There were no differences in knowledge regarding gender. When it comes to study programs, the students of Medical laboratory diagnostics had more knowledge about nosocomial infections. Furthermore, the level of students' knowledge was proportional to the amount of classes on the prevention of nosocomial infections attended. In other words, those who have attended more classes, showed better knowledge in the studied area. Conclusion: Although the students showed an adequate level of knowledge about the prevention of nosocomial infections, there is an insufficient level of knowledge about the proper implementation of hand hygiene, more precisely, about the indications for the use of alcohol-based hand rubs (antiseptics