Background:Hepatitis C Virus (HCV) is a major public health problem worldwide. About 130- 200 million people are infected with HCV worldwide leading to 500,000 deaths annually (WHO 2014). Healthcare workers (HCWs) have played an important role in the transmission of HCV infection, either as victims or as sources of infection.Objectives: To determine the prevalence of HCV, antibodies (Abs) RNA and genotypes among the female HCWs in Baghdad and to identify whether HCWs were infective or only infected.Subjects and Methods:A cross-sectional study involving 1001 women attending 17 health care centres in Baghdad, Iraq, was carried out. Information on type and duration of their occupation was obtained. HCV Abs (anti-HCV) were tested using a third generation enzyme immunoassay (EIA-3) and immunoblot assay (Lia Tek-111). Molecular analysis using RT-PCR and DNA enzyme immunoassay (DEIA) for HCV-RNA and genotype detections were carried out for 63 serum samples.Results:Only 160/1001 (15.98%) were HCWs. Anti-HCV and HCV- RNA seroprevalence were significantly higher (6.37%, p=0.0057, 88.83%, p= 0.011 respectively) among HCWs than non HCWs. HCWs were at a significantly higher risk of exposure to HCV infection (OR=2.75, 95% C.I. =1.31-5.79). There was no significant association between HCV genotypes and the HCWs. HCV-4 showed higher expression (62.5%) among HCWs.Conclusion:Female HCWs were infective and infected with HCV, thus there is a need for medical equipment to be sterilized and cleaned thoroughly.