Background Transfusion-transmitted infections threaten the safety of patients who require a blood transfusion, which in turn imposes serious challenges for the availability of safe blood products that are still affordable in healthcare systems with limited resources. Aim of the study The aim of the study was to determine the seroprevalence of transfusion-transmitted infections in blood donors at TMU Blood Bank and access the level of safety. Method A laboratory-based descriptive, cross-sectional study was conducted from June 2022 to Dec 2022. A qualitative study was carried out to access serological analysis of replacement and voluntary donors at TMU Blood Bank. The total number of donors screened during this period was 776. The donors were requested to fill out a detailed questionnaire and were screened for HIV, HBV, HCV, and Syphilis with a rapid immunechromatographic kit. The results were expressed in tables and figures. Results Out of 776 donors, 676(98.7%) were replacement and only 10(1.29%) were voluntary donors. Male donors were predominant constituting 676(87.12%) and only 100 (12.88%) female donors. The seroprevalence of TTIs in the present study was 1.42%. Syphilis (0.77%) was the most prevalent TTI followed by HBV (0.51%). A low prevalence of HCV (0.13%) was found and HIV was not detected in any donors in the present study. The highest prevalence of TTIs was observed in the young age group of 18-25 years followed by 26-40 years.