Introduction: Transfusion Transmissible Infections (TTIs) such as Human Immunodeficiency Virus (HIV), Hepatitis B virus (HBV), Hepatitis C Virus (HCV) and syphilis pose a threat to blood safety in a developing country like India. Aim: To determine the prevalence of TTIs among healthy donors in Andaman and Nicobar Islands, India, and to further help in determination of the population subset to be focused on for enhancing blood donor pool. Materials and Methods: This retrospective cross- sectional study was conducted on all voluntary and replacement donors in a tertiary health care centre in Andaman and Nicobar Islands for a period of three years. Donor blood was screened for HIV, HBV, HCV, Syphilis and malarial parasite using rapid kits and ELISA. Results: Out of 12,118 donors, 97.01% were males and 2.99% were females. 9409 (77.64 %) were voluntary donors and 2709 (22.35 %) were replacement donors. Majority of the screened donors belonged to 25 to 40 age group. Out of 12,118 donors, 265 (2.18 %) were seropositive. Among them, 8 (0.066%) were HIV positive, 15 (0.124%) HCV positive, 128 (1.056%) HBsAg positive, 30 (0.247%) Syphilis positive and 84 (0.693%) were positive for malarial antigen. Yearly prevalence of seropositivity was 2.370%, 2.113% and 2.082% for three consecutive years. Conclusion: With high prevalence of TTIs among healthy donors, safe blood transfusion is still a challenge in developing countries. This continues to place unacceptable burden on health sector. There is a need for encouragement of voluntary blood donation, strict donor selection criteria and sensitive screening tests to minimize TTIs in Indian setting.