Background: Transfusion transmissible infections (TTIs) are one of the major adverse effects of blood transfusion. Serological testing can reduce, but not eliminate the occurrences of TTIs. Study of TTIs amongst blood donors could help to find safe blood for the patients. Aims: This study was conducted to evaluate the prevalence of markers of Hepatitis B virus (HBV), Hepatitis C virus (HCV), Human Immunodeficiency Virus (HIV), Syphilis and Malaria among Voluntary Blood Donors and Replacement Blood Donors in a tertiary care centre. Materials and Methods: A retrospective study was conducted at The State of the art Model blood bank, Pt. Jawaharlal Nehru Memorial Medical College (JNMMC) and Dr. B. R. Ambedkar Memorial Hospital, Raipur, Chhattisgarh. This study included all the blood donors both from in–hospital donations and outdoor camps donations (including Voluntary and Replacement donors) from January 2011 to December 2018. Enzyme–linked immunosorbent assay (ELISA) test was done for screening of HBV, HCV and HIV; Rapid Plasma Reagin (RPR) test was done for syphilis and Card Test for Malaria Antigen. Results: A total of 1,08,577 donors were screened over a period of eight years. The seroprevalence of HIV, HBV, HCV, syphilis and malaria was found to be 0.12%, 0.77%, 0.04%, 0.22% and 0.03% respectively. Overall seroprevalence of reactive TTIs was 1.19%, which was maximum in the year 2013 (2.28%) after that there was a gradual decrease in the prevalence rate of TTIs in blood donors over the years. Infections were more common among Replacement Donors as compared to Voluntary Donors. Conclusion: The increase in public awareness regarding Voluntary blood donation, meticulous donor screening, counselling and use of highly sensitive tests can help in reducing the risk of TTIs. Prevalence of HIV infection is decreasing in the blood donors. The HBV infection still remains a menace to be tackled. HCV, Syphilis and malaria maintain a low rate of positivity. Methods to ensure a safe blood supply should be encouraged.