Background: In our healthcare setting transmission of hepatitis B is a big challenge to handle for both patients and healthcare providers especially those in frequent contact with blood. Objectives: To determine Hepatitis B knowledge, attitude, and vaccination status among nurses of Tamale Central Hospital. Methods: This study was conducted using a descriptive cross-sectional survey. Data analysis was done with SPSS version 20. Bivariate analysis was done using chi-square to determine the relationships. Results: The study recorded a response rate of 92.5%. Majority (59.6%) had high, 36.4% had moderate and 4.0% had low knowledge hepatitis B. Hepatitis knowledge level was associated with: age X2 (4) = 17.789, P= 0.001, sex, X2(2) = 13.203, P = 0.001, educational level, X2(6) = 17.552, P = 0.007, nursing category, X2(4) = 19.226, P = 0.001, and duration of nursing practice X2(2) = 19.492, P ≤ 0.001. About 42.9% had positive attitude toward hepatitis B prevention and attitude level towards hepatitis B was associated with: marital status X2(1) = 11.090, P = 0.001, residential address X2 (2) = 11.411, P = 0.003 and duration of nursing practice X2(1) = 4.769, P = 0.029. About 84.8% of nurses started vaccination against hepatitis B and 77.3% completed at least three doses of the vaccination. The only factor with a statistically significant relationship with vaccination completeness status was respondent marital status X2(1) = 8.063, P = 0.005. Conclusion: Nurses' knowledge of hepatitis B was very good. And more than half of the nurses had a negative attitude towards hepatitis B prevention. In terms of vaccination more than three-fourth of them started and completed three doses of hepatitis B vaccination.