Background: Eliminating high-risk individuals has a special role in ensuring blood safety. Due to epidemiological, demographic, and even cultural changes in each country, this process should be continuously evaluated and reviewed, if necessary. Objectives: This study aimed to evaluate the impact of the current donor selection procedure on blood safety in Iran. Methods: A total of 2,525 high-risk deferred donors who were referred between 2018 and 2019 were evaluated regarding hepatitis B surface antigen, hepatitis C virus antibody, and human immunodeficiency virus antigen and antibody. All repeatedly reactive samples were evaluated by confirmatory tests. Characteristics' parameters, donor status, and TTI marker rates of the participants and 1,315,871 eligible donors in the indicated period obtained from the national database on blood donors, were compared. Data were analyzed using SPSS version 24.0. Results: The prevalence of HBV, HCV, and HIV in 100,000 deferred donors was 1148, 515, and 119, respectively. This prevalence was 26, 28, and 33-times higher than the eligible donors, respectively. Unlike HBV, its prevalence among males was almost twice that of females among the deferred group. In the eligible group, females had a higher prevalence for HBV and HCV as compared to males. The HCV and HBV (6.7 and 4.3-fold) among deferred first-time donors had a significantly higher prevalence compared with the eligible first-time donors (P‐value < 001). Notably, the higher was the education degree, the lower was the prevalence of infection in both groups. Conclusions: Current deferral criteria and donor selection procedure in Iran are an opportunity to eliminate high-risk individuals from the blood donation.