Hepatitis C virus (HCV) infection is a major public health problem worldwide and remains a vital cause of chronic hepatitis. This study was aimed to detect the prevalence of HCV infection among Iraqi people. Four subjects of hemodialysis (HD) patients, thalassemia patients, blood donors and medical staff were enrolled in this study and evaluated for their serum anti-HCV-immunoglobulin G (IgG)-antibodies (Abs). The total frequency % of IgG anti-HCV Abs positivity was 3.2%, in which the highest frequency % was recorded among thalassemia patients followed by HD patients and then medical staff subjects, whereas the lowest frequency rate was reported within blood donors group. The frequencies of IgG anti-HCV Abs positivity were significantly elevated in males compared to females. For age groups, the results revealed higher infection rate of HCV among age group of 1-20 year followed by the age group of 21-40 year and then age group of 41-60 year, whereas the lowest rate of infection was recorded in age group >60 year. In conclusions, the prevalence rate of HCV infection among Iraqi people is similar to those in most of Asian and non-Asian studied populations and the infection rate was higher in males and inversely correlated with age of the patients. Blood transfusion, renal dialysis and health care workers (HCWs) were major sources of HCV infection. Thus, we recommend continuing surveillance of blood donors, HCWs and patients, in addition to HCV markers screening by molecular technique for the diagnosis of HCV during the window period in order to decrease the prevalence of HCV infection.