When talking about any health intervention, the first and indispensable step is hand hygiene. Throughout history, many infections and diseases have been associated with insufficient hand hygiene specifically. Improper or insufficient hand hygiene in healthcare workers leads to the transmission of microorganisms that are not part of the normal flora and easily spread among healthcare professionals and patients. The most effective way to reduce infection by transmitting microorganisms is hygienic alcohol rubbing and hand washing. This paper looks at hygiene indications that include the so-called five moments of hand hygiene and the ways in which the act of hygiene itself can be performed. The central part of the paper focuses on nursing students who are already familiar with the health system through their education and practical work, but also as future professionals who should leave the faculty with the knowledge and routine of hand hygiene. The aim of this paper is to survey the knowledge of undergraduate nursing students in the Republic of Croatia on proper hand hygiene. The research was conducted through an online survey that was distributed on social networks via the Google form in the period from September 7, 2020, to September 10, 2020. A total of 82 undergraduate nursing students from four counties participated in the survey. The survey questionnaire consisted of 53 questions with multiple-choice answers. The results were analyzed in two large groups, the knowledge, and attitudes of students. The analysis of the survey shows a satisfactory level of knowledge of students about the "Five Moments for Hand hygiene" method, as well as their positive attitudes towards the importance of hand hygiene. Students believe that hand hygiene is one of the most important things in health care, which indicates their knowledge of the topic, but also confirms the positive attitudes expressed. The results of the survey indicate the confirmation of both hypotheses set at the beginning of the paper.