The criterion of six or more mutations in the IRRDR (IRRDR ≥ 6) was identified as the most powerful viral genetic factor that independently predicted SVR (15).

In another study curried out on a patient cohort in Yamagata Prefecture, Japan, we proposed that polymorphism in the secondary structure of the N-terminal region of NS3 of HCV-1b influences virological responses to PEG-IFN/RBV therapy, and that virus grouping based on NS3 polymorphism can also be used to predict the outcome of the therapy (16). In the present study, we further analyzed the Yamagata cohort for a possible Gamma-secretase inhibitor relationship between heterogeneity of NS5A and the core regions of the HCV genome and virological responses to PEG-IFN/RBV therapy.

Fifty-seven patients who were chronically infected with HCV-1b, their diagnoses being based on detection of anti-HCV antibody and HCV RNA, and who had been seen at Yamagata University Hospital in Yamagata, Japan, were enrolled in the study. Their HCV subtypes were determined according to the method of Okamoto et al. (17). Patients were treated with PEG-IFNα-2b (Pegintron; Schering-Plough, Kenilworth, selleck compound NJ, USA) (1.5 μg per kilogram of body weight, once weekly, subcutaneously) and RBV (Rebetol; Schering-Plough) (600∼800 mg daily, orally), according to a standard treatment protocol for Japanese patients established by a Hepatitis Study Group of the Ministry of Health, Labor and Welfare, Japan. All patients received >80% of the scheduled doses of PEG-IFN and RBV. Serum samples were collected from the patients before treatment and at intervals of 4 weeks during the whole observation period (72 weeks), and tested for HCV RNA titers as reported previously (18). The study protocol was approved beforehand by Atorvastatin the Ethics Committee at Yamagata University Hospital, and written informed consent for study participation was obtained from

each patient prior to treatment. Also, the study protocol conforms to the provisions of the Declaration of Helsinki. Hepatitis C virus RNA was extracted from 140 μL of serum using a commercially available kit (QIAmp viral RNA kit; Qiagen, Tokyo, Japan). Amplification of full-length NS5A and the core regions of the HCV genome were performed as described elsewhere (11, 18, 19). The sequences of the amplified fragments of NS5A and core regions were determined by direct sequencing without subcloning. The aa sequences were deduced and aligned using GENETYX Win software version 7.0 (Genetyx, Tokyo, Japan). To evaluate the optimal threshold of the IRRDR and ISDR mutations for SVR prediction, we constructed an ROC curve and calculated the AUC, sensitivity and specificity (11). Statistical differences in treatment responses according to NS5A and core sequence heterogeneity were determined by the χ2 test.