Target sequences were FK228 mw amplified by means of polymerase chain reaction with specific primers. The nucleotide sequences of the amplicons were determined directly in both forward and reverse directions, using the ABI Prism BigDye Terminator cycle-sequencing ready reaction kit
on a fluorescent model 3100 DNA sequencer (Applied Biosystems). The amino acid sequences were deduced and aligned using Win software (version 7.0; Genetyx, Tokyo, Japan). Throughout this article, the amino acids are numbered according to the full-length genome sequence of isolate H77 (GenBank and DDBJ accession number AF009606). Patients infected with HCV genotype-1 received treatment once per week for 48 weeks with recombinant peginterferon alfa-2b (Peg-intron, Schering-Plough Pharmaceutical, Osaka, Japan), at a dosage of 1.5 μg/kg body weight, in combination with oral ribavirin (Rebetol, Schering-Plough Pharmaceutical) as previously reported. Those with HCV genotype-2 received the same treatment regimen for 24 weeks. Ribavirin was administered orally in a weight-adjusted dose given in two doses. The ribavirin dose was 15 mg/kg per day in patients
weighing ≤40 kg, 600 mg/day in patients weighing 40–60 kg, 800 mg/day in patients weighing 60–80 kg, and 1000 mg/day in patients weighing >80 kg. Blood tests and physical examinations were performed at 2, 4, 6, and 8 weeks after initiation of therapy, then every 4 weeks selleck inhibitor until
the end of treatment. After finishing treatment, blood tests and examinations were done at 12 and 24 weeks during the follow-up period. Adolescents among our patients were instructed to use birth control find more during the treatment and for 6 months after the end of treatment. Differences in mean values and the frequency of patients’ characteristics between groups were compared using the Mann–Whitney U-test and the Fisher’s exact test, respectively. All statistical analyses were performed using the SPSS V.12.0 statistical package, based on two-sided hypotheses tested with a significance level of P < 0.05. The study protocol complied with the ethical guidelines of the Declaration of Helsinki of 1975 (2004 revision) and was approved by the Ethics Committee of Osaka General Medical Center as well as by the respective organizations of each participating institute. The treatment response was evaluated 24 weeks after the completion of the prescheduled treatment period in 30 patients. The subjects’ average age was 10.9 years (range 3 to 17 years), the male-to-female ratio 16:14, transmission route was maternal infection in 26 patients and four infected from a blood transfusion, and the genotype-1: genotype-2 patient ratio was 16:14 (Table 1). The 30 patients in the study consisted of 17 children (3–11 years) and 13 adolescences (12–17 years).