Introduction Vaccination or active immunization is a procedure in which certain substances are introduced into the human body, and by means of which the immune system is informed about the causative agent with which it has not yet come into contact. The aim of this research was to examine the knowledge and attitudes of undergraduate professional nursing students at the Faculty of Health Studies in Rijeka about vaccination. Materials and methods The students of the undergraduate professional study of nursing at the Faculty of Health Studies in Rijeka participated in the research. The questionnaire was administered via Google Forms, and 100 respondents filled it out. Respondents of both sexes, divided into 4 age groups from 18 years old to over 50 years old, were included in the research. Results Out of 100 respondents who participated in the research, 93 respondents (93%) were female and 7 respondents were male (7%). The results showed that there is no significant statistical difference in knowledge (regular - Me (IQR)) and attitudes (regular - Me (IQR) 7 (6 – 8); extraordinary - Me (IQR) 7 (5 – 8)) attitudes (regular - Me (IQR) 59 (56 – 65); part-time - Me (IQR) 60 (55 – 65)) of students depending on their student status (full-time or part-time). The results showed that there were no significant statistical differences in the students' knowledge according to their status (χ2 test; p > 0.05), and that there is no significant statistical difference in attitudes towards study status (Mann Whitney test; p > 0.05). Conclusion The topic of vaccination among students encourages discussion, and is extremely important because nurses are the first in contact with patients. During this research, a significant statistical difference in students' knowledge was demonstrated depending on theirs high school education. The results showed that students who completed secondary medical school have significantly better knowledge than students who completed gymnasium. It has also been proven that there is a connection between students' knowledge and attitudes. The results show that the better the student's knowledge about vaccination the more positive the attitudes.