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目的 探讨丙型肝炎病毒 1b型 (HCV 1b)NS5A区基因的复杂性与干扰素α(IFN α)治疗效果之间的关系。方法 以 13例接受IFN α治疗的HCV 1b感染患者为研究对象 ,治疗前血清提取的RNA采用逆转录套式PCR (RT nested PCR)扩增HCVNS5A区基因 ,PCR产物纯化后与T载体连接并克隆入大肠杆菌 ,随机挑选 30个阳性克隆菌落 ,每一克隆菌落再进行PCR扩增 ,30个克隆的PCR产物在同一聚丙烯酰胺凝胶上采用单链构象多态性分析 (SSCP)分析和异源性双体分析 (HD)筛选HCVNS5A区克隆型 ,克隆型的多少反映HCVNS5A区基因的复杂程度。结果 完全应答组平均 6 .3种克隆型 ;部分应答组平均 8.6种克隆型 ;无应答组平均 15 .8种克隆型。结论 HCVNS5A区基因的复杂程度与IFN α的治疗效果呈负相关。
Objective To investigate the relationship between the complexity of the NS5A gene of hepatitis C virus type 1b (HCV 1b) and the therapeutic effect of interferon α (IFN α). Methods Thirteen patients with HCV 1b infection treated with IFNαwere enrolled. HCV RNA extracted from the serum before treatment was used to amplify HCV NS5A gene by reverse nested PCR. The PCR product was purified and ligated with T vector and cloned Enter into Escherichia coli, 30 positive clones were randomly selected, and each clone was further amplified by PCR. The PCR products of 30 clones were analyzed by single strand conformation polymorphism (SSCP) on the same polyacrylamide gel Sourced dual-body analysis (HD) screened HCVNS5A region clone, clone number reflects the HCVNS5A region of the complexity of the gene. RESULTS: An average of 6 clonotypes were found in the complete response group, an average of 8.6 clonotypes in the partial response group, and an average of 15.8 clonotypes in the no-response group. Conclusion The gene complexity of HCV NS5A region is negatively correlated with the therapeutic effect of IFNα.