Aim: To determine the current sero-prevalence of HCV among blood donors in Sana'a city: Yemen. Study Design: A descriptive cross sectional study. Place and Duration of Study: This study was conducted on 3606 blood donors at the National Blood Transfusion and Research Centre in Sana'a, Yemen, during the period from October 2016 to January 2017. Methodology: A descriptive cross-sectional study was conducted to estimate the prevalence of anti-HCV antibody using enzyme immunoassay (EIA) technique at the National Blood Transfusion and Research Centre in Sana'a Yemen during the period from October 2016 to January 2017. Results: A total of 3606 blood donors were enrolled in this study, 1.6% of them were anti-HCV positive. Distributed as the following, 1.1%, 2.8%, 2.2%, 2.9%, 3.4%, 2.4% and 1.7% of anti-HCV antibody was among blood donors resident the capital city of Sana'a, Sana'a rural area, Taiz, Ibb, Dahmar, Amran and Almahwet governorate respectively. The prevalence rate of HCV was higher among Handicraft (1.8%). High HCV seropositivity were detected among the age group 36-45, the difference was found statistically insignificance. Conclusion: Low percentage of HCV sero-prevalence was detected among our blood donors. Specific strategies for the screening of blood donors should be followed in the blood bank centers in Yemen.