Hepatitis C virus infection is a major public health problem that mainly affects the liver, causing liver cirrhosis, hepatocellular carcinoma, and liver failure, if it was untreated or treated late in patients with kidney transplantation and hemodialysis patients leading to a serious problem in this population. The aim of this study to determine the infection rate and genotypes of the hepatitis C virus among hemodialysis patients in Diyala governorate-Iraq. A cross-sectional study was performed on hemodialysis patients who were admitted to the Baqubah Teaching Hospital - Ibin Sina Dialysis Center and Khanaqin General Hospital, during the period from 16th September 2020 until 16th December 2020. Plasma was separated from 5 ml of a blood sample obtained from 306 hemodialysis patients (177 were males and 129 were females), their ages ranged from 10-85 years, then use for serological and molecular detection after RNA extraction from those HCV antibody seropositive hemodialysis patients. Furthermore, genotyping of the hepatitis C virus -RNA positive samples was done by one step using the real-time polymerase chain reaction technique. The results showed the infection rate of HCV in hemodialysis patients in Diyala governorate was 7.8% (24 out of 306), no significant differences were noticed between males and females, the age between 61-70 years old were more susceptible to infection with highly significant differences (P=0.0087), the blood transfusion was most risk factors to infection with highly significant differences (P=0.0054). Genotyping was performed in all patients who were HCV-RNA positive. Genotypes 4, 1a, 1b, and 2 were found in (33.33%), (29.2%), (29.2%), and (8.3%) respectively. This study concluded the infection of HCV among hemodialysis patients in the Diyala governorate was moderate and genotype 4 was more predominant followed by genotype 1 (a, b) and more common in treated patients, no significant differences were noticed between males and females, the ages between 61-70 years were most susceptible to infection, and blood transfusion is the most risk factors for transmission of infection in this population

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