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• All positive viral markers in blood donors were identified in new donors. • Pre-donation laboratory screening can reduce the risk of TTI transmission by identifying infected new donors at the pre-donation phase. • Pre-donation screening program can lead to donor loss. This donor loss resulted the more effort to retain blood donors to compensate the donor loss. The risk of transfusion-transmissible infections (TTIs) remains a concern in transfusion medicine. Since the rate of infection among first-time blood donors is higher than repeated donors, strategies to enhance blood safety can focus on new donors. The aim of the study was to investigate the effect of pre-donation viral screening of new donors on blood safety. The pre-donation screening of new donors was implemented in the Kurdistan blood center. In this program, new donors who met the blood donation criteria were informed about the program and only a blood sample was donated for HBs Ag, HCV Ab, and HIV Ab testing. New donors with negative results were invited to donate blood after 12 weeks. A unit of blood was collected from eligible returned donors. Laboratory tests were performed again using the same methods. Finally, the prevalence of confirmed positive TTI results among donated blood in Kurdistan blood center was compared before and after the establishment of program. During the study, 4,434 new donors were screened for viral markers. A total of 41 new donors (0.92%, 95% CI, 0.007–0.13) had repeatedly reactive results and infection was confirmed in blood sample of 24 donors (0.54%, 95% CI, 0.003–0.008). Overall, 56% of new donors returned for blood donation. Prevalence of confirmed TTIs markers in collected blood units was 0.27% and 0 before and after implementing program, respectively. This study indicated that Pre-donation screening can reduce the risk of TTI transmission by identifying infected donors at the pre-donation phase. [ABSTRACT FROM AUTHOR]