Background: Hepatitis B virus (HBV) infection is a global problem and >350 million HBV carriers in the world. Objectives: The aim was to assess awareness, practice of selected high risk health care workers (HCW) regarding risk for contracting hepatitis B and self-reported vaccination status. Materials and Methods: A cross-sectional study was conducted among 300 selected HCW of AGMC and GBP Hospital, TMC and Dr BRAM Teaching Hospital and Indira Gandhi Memorial Hospital, Agartala during Jan to March 2014. Results: About 72.1% respondents were females with overall mean age of 24.10 (standard deviation ±7.011). 67.5% females were within the age group of 18-23 years. Majority (70.4%) of the participants were nursing students and Hindu (92.5%) nuclear families (54.6%). Majority (69.3%) of them were aware that hepatitis B transmission was possible through unsafe sex, infected blood/body fluid, contaminated syringe, needle and scalpel, 19.6% knew through infected blood and body fluids, 5.7% knew through contaminated syringe, needle and scalpel, 1.8% knew through unsafe sex. 59.3% had a history of contact with known hepatitis B case. 62.2% were vaccinated with three doses of hepatitis B vaccine. Majority of the participants had exposed to hepatitis B positive case while at work (P = 0.001). The exposed persons with known hepatitis B cases have consulted doctor, vaccinated and treated with medicines (P = 0.002); used needle destroyer (P = 0.012); vaccinated with 3 doses of hepatitis B vaccine (P = 0.001); and used sterile gloves while performing work (P = 0.000), especially while dealing with blood and body fluid. Conclusion: In spite of having good knowledge, the way they practice for prevention of hepatitis B infections were inadequate and need further improvement.