Background: Hepatitis C virus (HCV) infection is a worldwide public health problem where its seroprevalence had an estimated 2.8% increase over the last decade. Objective: To assess the changes in HCV infection rates among Egyptian blood donors, 12 years after implementation of anti-HCV treatment program. Patients and methods: This was a prospective cross-sectional study conducted at Al-Azhar University Hospitals, and Mansoura New General Hospital. A total of 2000 consecutive blood donors were tested for anti-HCV antibodies. All donors filled blood donor questionnaire. Anti-hcv positive donors were subjected to hepatitis C ribonucleic acid (HCVRNA) testing by real-time polymerase chain reaction (PCR). Results: The majority of donors (n=1992; 99.6%) were males. A total of 1515 (75.75%) donors were living in rural areas, while 485 (24.25%) were living in urban areas. In addition, 862 (43.1%), 620 (31%) and 466 (23.3%) graduated from secondary, high and primary schools, respectively. Blood donors whom graduated from higher education were less likely to be anti-HCV-positive (P ≤ 0.05). Among 35 anti-HCV positive blood donors, 6 (17.15%) were HCVRNA positive. Blood donors with positive viremia were older (34.0± 5.29 years) than those with negative (29.66± 6.75 years) viremia. Intravenous drug user, previous needle stick injury, previous surgery, family history of viral hepatitis, previous hospitalization and history of anti-HCV treatment were identified as risk factors for anti-HCV positivity (P < 0.01). Out of the 2000 blood donors, only 5 (0.25 %) males with mean age of 30.6± 3.29 years were positive for HBs Ag. Lower education levels (P ≤0.05), marriage (P ≤0.05) and previous surgery, (P ≤0.01) were identified as risk factors for HBV infection. Conclusion: A significant reduction of HCV prevalence among Egyptian blood donors was detected in our survey. In addition, a lower rate of HBsAg among blood donors was also detected. Notably, the iatrogenic transmission of HBV and HCV still ongoing in Egypt.