Background: Human Papillomavirus is a common sexually transmitted infection, representing the main cause of genital warts and cervical cancer. The objective of this study was to evaluate basic knowledge and beliefs regarding HPV infection and HPV vaccine among undergraduate healthcare men and women students, as well as their attitudes towards HPV vaccine. Methods: Undergraduate women and men (nursing and midwifery curses) attending three Schools of Health Sciences located in Switzerland. A total of 427 women and 223 men have completed the web questionnaire, which included questions on their socio-demographic background and about basic knowledge and attitudes toward the HPV infection and vaccination. Results: Women undergraduate students have a better knowledge of HPV infection than their men counterparts, although there was a significant gap in knowledge of the disease’s mode of transmission and prevention. Among women, 72.6% of respondents reported having received at least one dose of HPV vaccines versus 31.4% for men respondents. Conclusion: The results of this study revealed a poor understanding among undergraduate healthcare men and women students about the HPV infection, its mode of transmission and its prevention. Our findings highlight the need to improve education on HPV for undergraduate healthcare students in order to increase the awareness of the disease.