Background <p>Hepatitis B virus is a blood borne disease that poses a significant public health problem and is classified as an occupational hazard for healthcare workers, including nursing students. Healthcare workers are four times more likely to contract HBV than the general population. Therefore, a thorough understanding of HBV is crucial for nursing students. This study aimed to assess the level of knowledge about HBV infection among nursing students at Jiblah University of Medical and Health Sciences.</p> Methods <p>A descriptive cross-sectional study was conducted. This study included all 94 students enrolled in the Faculty of Nursing. A standardized, pre-tested questionnaire consisting of three sections was used: students’ sociodemographic data, their knowledge of hepatitis B virus infection, and their knowledge of hepatitis B treatment and prevention. The collected data were analyzed using SPSS version 26. The chi-squared test value was set at 0.05. A p-value less than 0.05 is considered statistically significant.</p> Results <p>The results showed that the mean age of the participants was 22.15 ± 2.32 years, and that 57.47% were female. Overall, 73% of students had a good knowledge of hepatitis B virus. Forty-six students (48.9%) in the study group were vaccinated against HBV. A lack of knowledge was found regarding HBV transmission methods. Students’ knowledge of HBV infection was associated with gender and year of study.</p> Conclusions <p>The study indicated that students had a good knowledge of hepatitis B virus infection. There were statistically significant correlations between students’ knowledge of hepatitis B virus infection and their gender and year of study.</p>