The objectives of this study were to document the background prevalence and incidence of HCV infection among HCWs in Ain Shams University Hospitals in Cairo and analyse the risk factors for HCV infection. A cross-sectional survey was conducted in 2008 among 1770 HCWs. Anti-HCV prevalence was age-standardized using the Cairo population. A prospective cohort was followed for a period of 18 months to estimate HCV incidence. The crude anti-HCV prevalence was 8.0% and the age-standardized seroprevalence was 8.1%. Risk factors independently associated with HCV seropositivity were: age, manual worker, history of blood transfusions and history of parenteral anti-schistosomiasis treatment. The estimated incidence of HCV infection was 7.3 per 1000 person-years. HCWs in this setting had a similar high HCV seroprevalence as the general population of greater Cairo.