Background: Advanced techniques like Chemilumenescence and NAT are being used to eliminate the risk of TTIs. Safety measures starts right from the donor screening and donor history questionairre by which individuals who are infected with pathogens having acute symptoms are likely to self defer that will reduce the chance of TTIs among the recepient population during window period. Objectives: To determine the seroprevalence and trends of major transfusion transmissible infections in blood donors. Material and Methods: A cross-sectional retrospective study (2019-2022) was conducted in the Department of Blood Transfusion Medicine (Blood Centre), Government Medical college and associated hospital, Rajouri, Jammu and Kashmir, India. Serum samples were tested by enzyme-linked immunosorbent assays (ELISA) for the presence of hepatitis B surface antigen (HBsAg), and antibodies to human immunodeficiency virus (HIV-1/2), hepatitis C virus (HCV). Syphilis and Malaria screening was done by rapid card method. Results: A total of 5682 apparently healthy blood donors were tested. Out of which 4119 (72.49%) were replacement donors and 1563 (27.50%) were voluntary donors. The majority of them were males 5648(99.4%) and aged between 31 -40 years(46.4%). A total of 49 (0.86%) donors had serological evidence of infection with at least one pathogen. Among them 4(8.16%) were voluntary donors donated in outdoor camps out of which 2 were females having HCV positive.3 had responded to the call and after confirmation, were referred to the concerned department for treatment. 45(91.8%) were replacement donors, 10 had responded to the calls, 4 were confirmed and were referred for treatment rest all were non responders. Bivariate analysis of gender- wise prevalence of HCV was statistically significant (p<0.001) but HBV has no statistical significance(p=1). Year-wise seroprevalence and trend of TTIs shows a high significance(p<0.001). The positivity rates of HBV, HCV and HIV, syphilis and malaria were 17(0.29%), 32(0.56%) and 0%, 0% and 0% respectively. Trends for TTIs (Hepatitis) showed a significant increase for the year 2020 (0.18%) to 2022(1.58%) with the increase in the number of donations. Conclusion: A significantly high prevalence of TTIs was detected and it remains to be the greatest threat to the safety of blood. Chemilumenescence and NAT testing should be adopted to detect the virus within shortest possible incubation time.