Blood transfusion services are a vital and integral part of modern healthcare services. However, the risk of transfusion transmittable infections (TTI) has been a major handicap. Therefore, this study was aimed at determining the prevalence of hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus (HIV) among blood donors. A retrospective study was conducted to collect data about the blood donors who consecutively donated blood from October 2011 to 2014. A three-year retrospective study was conducted in Mekelle Blood Bank. A data abstraction format was used to collect the sociodemographic and clinical data, and the prevalence of HBV, HCV, and HIV was determined. Data were analyzed using STATA version 10 analytical software. P value less than 0.05 was considered significant in all the analyses. A total of 10 728 blood donors, median (interquartile range) of age 30 (23-45) years and 3750 (34.9%) males were enrolled in this study. Of the participants 407(3.79%), 143(1.33%), and 111(1.03%) blood donors were positive for HBV, HCV, and HIV, respectively. HBV-HIV coinfections were found 10 (1.93%) blood donors, followed by HBV-HCV and HIV-HCV. A significant association between sex and marital status with HBV and HIV infection was found. However, significant association of HCV was observed among sex ( X  = 33.18, P < 0.001) and occupational ( X  = 84.33, P < 0.001). A significant percentage of HBV, HCV, and HIV among blood donors was observed. To select a donor and collect safe blood risk factors exposing blood donor should be studied, and community-based prevalence studies on TTI are also required.