In order to investigate variation in hepatitis C virus (HCV) nonstructural 5b (NS5b) nucleotide sequence and the relationship with interferon treatment, cDNA containing in this region were obtained by reverse transcription-polymerase chain reaction from the sera of 4 HCV RNA positive asymptomatic chronic carriers and 9 chronic hepatitis C patients. The cDNA were then directly analysed by polymerase-chain-reaction-directed sequencing. Compared with three published HCV type II/1b sequences (HCV-J, HC-C2 and HCV-BDS), the homology of HCV was lower in the patients with chronic hepatitis C (93.52% +/- 2.09% average), especially in nonresponders to interferon (92.70 +/- 1.90% average), than in asymptomatic HCV carriers (95.65 +/- 2.61%). In 5 patients, HCV RNA was eliminated by interferon treatment. In the remaining 4 interferon-treated HCV survived patients there were 9-48 nucleotide substitutions in HCV mutants out of 380 nucleotides, resulting 6-20 amino acids changes out of 126 amino acids. These results suggested that NS5b region variants play an important role in maintaining persistent infection and the evolution of NS5b is associated with the effect of interferon treatment.