Hepatitis B Virus (HBV), Hepatitis C Virus (HCV) and Human Immunodeficiency Virus I, II (HIV-I,II) are serious global and public health problem. Knowledge of the seroprevalence would enable early diagnosis, treatment of patients and also help in prevention of horizontal spread of infection by application of universal infection control measures. To determine the seroprevalence of HBV, HCV and HIV I, II infection in patients undergoing ocular surgery. A retrospective review of 560 patients who had undergone ocular surgery between Dec 2014 to June 2015 at Regional Institute of Ophthalmology, Patna, India, was done. Blood samples of patients were tested for the presence of Hepatitis B Surface Antigen (HBsAg) and anti-HCV antibodies by Enzyme Linked Immunosorbent Assay (ELISA) method, while HIV I, II was tested by ELISA and Western Blot as per National AIDS Control Organization (NACO), India guidelines. The number of patients with positive viral serology, age, sex ratio and nature of surgery were analysed. Out of 560 patients, 27 (4.8%) patients were found to be positive for viral serology. HBsAg was positive in 22 patients (3.92%). Out of these 14 patients (2.5%) were male while 8(1.4%) were females. Anti- HCV was positive in 3 (0.53%) patients. Out of these 2(.35%) were male and 1 (.17%) was female. HIV was positive in 2 patients (0.35%), both of whom were males. Keeping in view the number of cases of seropositivity in our study and various studies showing the presence of these viruses in aqueous and tears, we suggest that screening for viral markers should be made mandatory before ocular surgery so that early detection and treatment of patients can be done and risk of horizontal spread is minimized.