Objective: The present study aimed to assess the knowledge, attitude, and practice (KAP) of 3 rd - and 4 th -year medical, dental, and pharmacy students about hepatitis B (HB) infection at a private medical university, Malaysia. Methods: A cross-sectional, questionnaire-based study was conducted among 482 medical, dental, and pharmacy students of 3 rd - and 4 th -year degree program of Asian Institute of Medicine, Science and Technology University to assess their KAP about HB infection using 34 prevalidated questions by convenient sampling method. A questionnaire was administered to the students, and their responses were measured at “yes” and “no” scale. Students' responses were entered in SPSS version 22, and quantitative analysis was performed using descriptive statistics and nonparametric tests at P < 0.05. Findings: The medical, dental, and pharmacy students had good knowledge and practice with positive attitude about HB infection. Mann–Whitney U-test determined a significant difference in knowledge ( P < 0.001) and practice ( P < 0.001) scores between medical and pharmacy, attitude ( P < 0.001) scores between medical and dental, and attitude ( P < 0.001) and practice ( P < 0.001) scores between pharmacy and dental students. Students' age was correlated with their attitude, practice, and KAP scores and family income with their knowledge, attitude, practice, and KAP scores. Conclusion: Although students' knowledge and practice were good with positive attitude, all the students did not participate in health education program, screening, and vaccination of hepatitis B virus (HBV) infection which makes them more vulnerable to occupational HBV infection. Hence, it is recommended to organize a regular health education program for the students on screening and vaccination against HBV to prevent its infection.