The prevalence of hepatitis C virus (HCV) antibodies, HCV infection, and genotypes was studied in a rural population of the Central African Republic. In five villages, blood samples were taken from all the inhabitants present during the survey, belonging to Pygmies (299) and to Bantu and Banda ethnic groups (247). Using a second-generation ELISA screening and confirmation by immunoblot assay for the detection of HCV antibodies, all the Pygmies were negative, whereas seven Bantus/Bandas, aged > 35 years and with no familial relationship, were positive, giving a prevalence of 2.8% in this ethnic group. Five samples were also PCR positive; all belonged to genotype 4, but with three new subtypes identified by phylogenic analysis. These results indicate the co-existence of different HCV subtypes and raise questions about the natural transmission of HCV in this secluded population.