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Background: Prevalence of hepatitis C virus and that of its main genotypes varies between the worlds geographic regions. The risk factors for infection with HCV include blood transfusion, tattoing and injecting drug use., Objectives: To examine the prevalence of HCV and determine its main genotypes among a cohort of drug users in Kenya., Design: A laboratory based study., Setting: Hepatitis research laboratory in the Centre for Virus Research at the Kenya Medical Research Institute, Nairobi., Subjects: Three hundred and fourteen male and 19 female intravenous and non-intravenous drug users aged between 15-55 years., Results: Seventy four (22.2%) out of 333 samples tested positive for anti-HCV. Sixty nine out of the 74 serum samples were assayed for HCV RNA and 38 (55.5%) were positive. The RNA positive samples were further subjected to sequencing and 19 (73%) of the samples were classified as genotype 1a, while seven (27%) samples were classified as genotype 4. Genotypes 2, 3, 5 and 6 were not identified in this study., Conclusions: These results demonstrate a high HCV infection prevalence among this cohort of drug users (22.2%) as compared to that of the general population, which is estimated to be 0.2-0.9%. The study also confirms the presence of at least two major genotypes among Kenyan drug users (genotypes 1 and 4).