summary. Nonstructural 5A (NS5A) and the second envelope (E2) proteins of hepatitis C virus (HCV) have the potential to block interferon (IFN)-induced RNA-dependent protein kinase (PKR) and may therefore interfere with the response to IFN therapy, but controversy still exists regarding the relevance of this. This study aimed to assess whether mutations in these regions correlated with the response to combination therapy, IFN and ribavirin. Pretreatment parameters were analysed in 57 HCV-1b patients who had received IFN-α2b (3 or 5 MU three times weekly) and ribavirin (800–1200 mg per day) for 24 weeks. The amino acid sequences of the NS5A and PKR-eIF2α phosphorylation homology domain (E2-PePHD) were deduced from the corresponding coding sequence, which were determinated by direct sequencing of the HCV genome amplified by the polymerase chain reaction. Twenty (36%) patients achieved a sustained virological response (SVR). The mean number of amino acid substitutions in the NS5A–PKR binding domain (2209–2274), interferon sensitivity-determining region (ISDR) (2209–2248), and E2-PePHD sequence (659–670) in patients with and without SVR were 4.53 ± 3.31 vs 2.83 ± 1.78 (P = 0.094), 2.45 ± 2.74 vs 1.03 ± 1.32 (P = 0.042) and 0.25 ± 0.70 vs 0.03 ± 0.17 (P = 0.109), respectively. Patients with a mutant-type (≥ 4) NS5A–ISDR had a higher rate of SVR (six of nine, 67%) than those with wild-type (five of 22, 23%) (P = 0.038). Stepwise multiple logistic regression analysis of the factors (age, gender, viral load, cirrhosis rate, IFN dosage and amino acid substitutions) revealed that the mutation in NS5A–ISDR (≥ 4 vs < 4) was the only independent variable of treatment outcome. Our study showed that NS5A–ISDR mutations were correlated with the SVR to combination therapy in chronic HCV-1b patients in Taiwan.