Hepatitis C virus (HCV) infection is a major global health problem due to high chronicity rates, occurrence of severe hepatic diseases, and absence of an accurate therapy and effective vaccine. It is well known that viral genome is highly variable and HCV has at least six genotypes, each of them containing a series of subtypes. HCV genotypes exhibit geographical and epidemiological distribution. Genotype identification is clinically important to decide the dosage and duration of treatment since different genotypes exhibit variable response to treatment. The aim of this study was to determine the HCV genotypes in chronic HCV patients who were followed-up in Antalya Research and Training Hospital, Turkey. Anti-HCV and HCV-RNA positive blood samples obtained from 148 chronic hepatitis C patients (67 female, 81 male; mean age: 50.5 +/- 10.8, age range: 17-73 years) who were admitted to Antalya Research and Training Hospital Microbiology Laboratory during January 2011-June 2013, were included in the study. Epidemiological data of the patients and HCV genotype results were evaluated retrospectively. Viral genotypes were determined by real-time (Rt) PCR assay (Abbott Molecular Diagnostic, USA). HCV genotype (Gt)-1 was detected in 119 (80.4%) of the patients, of them 15.9% (19/119) were identified as subtype la and 75.6% (90/119) were subtype 1b. The prevalence rates of Gt-2, -3, and -4 were found as 3.4% (n= 5), 11.5% (n= 17), and 2% (n= 3), respectively. Gt-6 was not detected in our patients. Mixed infection with HCV types was detected in four patients (2.7%) by Rt-PCR; of these three were detected as Gt-1 and one was Gt-2 by RFLP (Restriction Fragment Length Polymorphism) and sequencing. The high prevalence of Gt-3 (11.5%) obtained in this study was attributed to the determination of Gt-3 in seven of 13 foreign national subjects. Rt-PCR method used in this study is user independent, standardized, automated, rapid and reliable method, however in case of detection of mixed types, the samples should be confirmed by other methods. In conclusion, we reported that the majority of the chronic hepatitis C infected patients had Gt-1b, and Gt-3 exhibited the highest rate ever reported by other studies from Turkey.