Back ground: Hepatitis C Virus (HCV) infection is one of the leading causes of morbidity and mortality in under–developed countries like Yemen. According to an estimate, 2.0 % of Yemeni’s total population suffers from HCV infection which makes a total of 600,000 people. Hepatitis C has emerged as a big challenge for Yemen. Health care workers are at increased risk of being infected due to tremendous exposure. Objective: The purpose of this study was to find the prevalence of hepatitis C, its risk factors contributing for infection among health care Workers at Sana’a City. Methods: A cross sectional survey conducted among 345 health care workers in the main general hospitals and health centers in Sana’a City. Blood samples were collected, and HCV antibodies by both Enzyme Immunoassay (EIA) and Recombinant ImmunoBlot Assay (RIBA) were used for diagnostic purpose. Results: The prevalence rate of Hepatitis C virus among HCWs was 4.1%. Those who used needle cupping, needle stick injury, surgical operation, the rate was (8.7%), (4.9%) and (4.2%) respectively. Conclusion: The frequency of HCV among health care workers is high. Awareness programs, training and continuing education should be mandatory and establishment of a national policy roadmap to increase knowledge about hepatitis C prevention. It will decrease the incidence of hepatitis C infection among health care workers.