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Objectives: To establish a nationwide epidemiological profile of transfusion-transmittable infection (TTI) markers among seemingly healthy blood donors to update policies required to ensure blood safety. Methods:A nationwide, cross-sectional study was designed to examine donor demographics and TTI prevalence during 2020 using data provided by the Ministry of Health, Saudi Arabia.Results: Collectively, a total of 375,218 whole blood units were donated, of which 32,758 (8.7%) were Original Articleexcluded due to TTI-related risk. The exclusion was based on a positive nucleic acid amplification test (NAT) or seroreactivity to hepatitis B virus (HBV), hepatitis C virus (HCV), human immunodeficiency virus (HIV), human T-cell lymphotropic virus (HTLV-I/II), syphilis, or malaria. Notably, the central (37.6%) and southern (33%) regions were the epicenters of TTI-reactive blood donors. Hepatitis B virus markers accounted for 85.7% and were the overall most prevalent of TTI-positive donations, followed by HCV at 5.9% and syphilis at 5.6%. In particular, anti-HBc and HBsAg were most prevalent in the south, while HBV NAT was highest in the center. Conclusion:Hepatitis B virus, HCV, and syphilis carry the greatest risk of TTI in Saudi Arabia. Including HBsAg screening is a necessary precautious measure to maintain blood safety.