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Background: Hepatitis C Virus remains a significant risk for both patients and medical staff in hemodialysis centres. The current study aims to determine the infection rate of hepatitis C virus, and risk factors associated with hepatitis C virus in hemodialysis patients in Diyala Governorate\ Iraq. Methods: A cross-sectional study was based on the processing of blood samples from 306 hemodialysis patients with routine hemodialysis at the Dialysis Centres in the Baqubah Teaching Hospital, and Khanqin General Hospital, were collected during the period 16thSeptember 2020 till 16thDecember 2020, after preparation of samples, enzyme-linked immunosorbent assay test was performed to detect on hepatitis C virus -antibody (Immunoglobulin G), then the seropositivity samples confirmed by real-time polymerase chain reaction technique. Results: The rate of hepatitis C virus infection among hemodialysis patients in Diyala Governorate was 7.8% (24/306), and there were no differences between males and females (12 cases) for each case. Age between 61-70 years was the most likely 9(37.5%). There were no differences between the accommodation of patients in Baqubah or around Baqubah (50%) for each one, and 22 married patients (91.70%) more than unmarried 2(8.30%). Patients with primary education were more frequent 9 (37.50%),hemodialysis duration ranging from 1-4 years showed more frequent than other periods 21 (87.5%). Conclusion: The rate of hepatitis C virus infection was moderate among hemodialysis patients in Diyala Governorate and there are no differences between gender and residence and the infection increases with the duration of dialysis.