BACKGROUND Hepatitis B infection is a major global health problem. The seroprevalence of Hepatitis B virus infection among general population attending at a tertiary care hospital is useful in assessing the magnitude of HBV infection and helps in devising preventive measures. The co-infection of Hepatitis B or Hepatitis C virus with HIV accelerates disease progression and may complicate the management of patients infected with HIV. This study was designed to determine hepatitis B virus infection among patients attending tertiary care centre in NorthEast India. MATERIALS & METHODS A total of 11399 serum samples, collected from patients attending outpatient or inpatient departments and ART centre, were screened for HBsAg using 3 rd generation ELISA (Qualisa HBsAg, Qualpro Diagnostics, Goa, India) in this retrospective study. RESULTS Seroprevalence of HBsAg was 1.23%. Seropositivity was seen more in males (2.2%) as compared to females (0.89%). Among males, highest occurrence was found in the age group of 21-30 (3.29%) years followed by 31-40 (2.94%) years and 41-50 (2.37%) years. Among females, highest prevalence was observed in age group of 51-60 (1.39%) years followed by 61-70 (1.15%) years and 41-50 (1.13%) years. CONCLUSION Present study showed that the overall rate of HBsAg positivity in our center was 1.23%. Males and younger age group were found to be more affected probably due to horizontal transmission.