Hepatitis C virus (HCV) is an RNA virus that can cause liver inflammation (hepatitis) and has the potential to become chronic and can progress to liver cirrhosis and hepatocellular carcinoma. Detection of HCV RNA infection, and genotype/subtype of HCV was performed on 70 blood sera of patients at the Hepatology Outpatient Clinic of Dr Soetomo General Hospital, Surabaya, Indonesia. Detection of HCV infection was carried out by Anti-HCV determination using enzyme immunoassay (EIA) technique, detection of HCV RNA by Reverse Transcription Polymerase Chain Reaction (RT-PCR) technique based on genome regions NS5b and 5'UTR, followed by electrophoresis with agarose gel. In positive PCR results, HCV genotype/subtype was determined by direct sequencing method using ABI 310 sequencer and sequencing results were analyzed by comparing the products with previously published HCV nucleotides. Sera were obtained from 41 (58.6%) male and 29 (41.4%) female patients. Anti-HCV was found positive in 17/70 (24.29%) patients and 16/17 (94.1%) was proved to contain HCV RNA when determined by RT-PCR technique. Patients with positive HCV RNA have the potential to transmit HCV infection. From the genotype/subtype analysis of sequencing results we obtained 2/16 (12,5%), 3/16 (18,75%) and 6/16 (37,5%), 1/16 (6,25%), 1/16 (6, 25%), 2/16 (12,5%), 1/16 (6,25%) HCV genotypes 1, 2, and HCV subtypes 1b, 1c, 2a, 3a, 3k respectively.Conclusion: In patients who went to the Hepatology Outpatient Clinic, Dr. Soetomo General Hospital Surabaya, we found positive Anti-HCV was 24,29%. In 94,1% of patients with positive Anti-HCV, HCV RNA was still detected and HCV genotype 1 with subtype 1b were still dominant HCV subtypes
