Background: Human Papillomavirus (HPV) is one of the most common viral infections worldwide, comprising high-risk types associated with cancers. The introduction of HPV vaccines has revolutionized prevention. The objective of this study was to assess knowledge and attitude towards Human papillomavirus infection and evaluate perception of Human papillomavirus vaccine among university students. Methods: A cross sectional multinational study was conducted on 543 students both male and female students from July 2024 to January 2025, using an online survey distributed in English and Arabic formed of 4 parts Sociodemographic part, knowledge classified in to 3 section (about HPV infection, cervical cancer and importance of HPV vaccine), Attitude section includes 9 questions about University student’s attitudes and beliefs and Perception towards Human papilloma virus vaccine section includes 4 sections (perceived susceptibility, Severity, benefit and barrier). Results: There were 63.5% of participants with inadequate knowledge, 57.1% with a bad attitude, and 53.6% with a low perception. Positive correlation between attitude and (knowledge& perception) (r=0.37 & P<0.001) (r=0.52, P<0.001) respectively, there was a statistically significant decrease in total knowledge level among theoretical students compared to scientific students. (P <0.001). Additionally, there was a statistically significant increase in the frequency of bad attitudes among theoretical students compared to students from scientific faculties (0.04, respectively). While in perception, there was a statistically significant increase in perceived susceptibility, perceived barriers, and total perception among theoretical students compared to scientific faculty students. (P=0.02, <0.001 & <0.001 respectively). Conclusion: High prevalence of inadequate knowledge, negative attitudes, and low vaccine perception toward HPV among university students, particularly among theoretical faculties and specific demographic groups. Targeted educational strategies are essential to enhance knowledge and vaccine acceptance.