Background: Hepatitis B is a well-documented occupational hazard for health care workers, including both doctor and paramedical personals. Vaccination is the best way by which one can arm oneself against Hepatitis B virus (HBV) infection. The present study was conducted to assess awareness, attitude and vaccination status of staff nurses of a teaching hospital in north India. Materials and Method: All the staff nurses (170) who gave consent to participate in the study were enrolled in the study. A self-structured close ended questionnaire comprising of three parts was used to collect information. First part comprised of questions on demography, second part assessed subjects’ awareness regarding HBV infection and third part comprised of questions on immunization status. Chi-square test and multiple linear regression analysis were used for statistical analysis. Results: The mean age of the study subjects was 24.9 ± 6.8 years. Awareness regarding HBV infection was present among 94.7% (161) of subjects. Universal Precautions were followed by 81.8% (139) of subjects. Only 18.8% (32) subjects were following correct method of disposal of Biomedical Waste (BMW). The odds of getting immunized were 4.46 times greater in subjects who had awareness of Hepatitis B infection than those who did not. Conclusion: Awareness regarding HBV injection and its coverage was high among the subjects. Very few subjects were aware regarding correct route of transmission emphasizing the need for effective intervention programmes designed to increase knowledge about HBV infection.