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目的 探讨上海地区丙型肝炎病毒 (HCV) 1b亚型慢性感染者的血清HCV非结构基因5A(NS5A)与干扰素 (IFN)疗效的关系。方法 收集上海地区 2 4例HCV1b慢性感染者在干扰素治疗前后及随访过程中的血清标本 ,定量检测治疗前血清HCVRNA ,用逆转录 聚合酶链反应方法扩增NS5A的干扰素敏感决定区 (ISDR)基因并进行测序和分析。另扩增干扰素应答类型不同的 3例患者治疗前后共 5株HCV病毒的NS5A全长序列 ,测序后作种系发生树分析及蛋白二级结构预测。结果 治疗前血清HCVRNA的定量结果显示 ,持续应答组的病毒滴度 (平均滴度 4 50× 1 0 4copies ml)明显低于复发组和无应答组 (平均滴度 1 82× 1 0 7copies ml)。 2 4例慢性丙型肝炎患者干扰素治疗前血清HCV的ISDR氨基酸序列与抗干扰素的HCV J株比较 ,1 3例为野生型 ,1 1例为中间型 ,无突变型。 6例完全应答者 3例感染的是野生型株 ,另 3例感染的是中间型病毒株。 5株HCV病毒的NS5A全长序列种系发生树显示 ,3种不同应答类型株在种系发生上分属 3个组别 ,无应答株与抗干扰素的HCV J株关系相近被归为 1组。蛋白质二级结构预测显示 ,上述病毒株NS5A蛋白在二级结构方面基本相似 ,仅在 2 2 55～ 2 2 89范围内有明显不同 ,这一区域与PKR结合域部分重叠。结论 HCVNS5A基因?
Objective To investigate the relationship between the serum HCV non-structural gene 5A (NS5A) and the efficacy of interferon (IFN) in patients with chronic hepatitis C virus (HCV) 1b subtype infection in Shanghai. Methods Serum samples from 24 chronic HCVb infected patients in Shanghai before and after interferon treatment and during follow-up were collected. Pretreatment serum HCVRNA was detected by quantitative polymerase chain reaction (RT-PCR). The interferon sensitive region ) Gene and sequenced and analyzed. In addition, five NS5A full-length sequences of HCV virus before and after treatment were also amplified in 3 patients with different types of interferon responses. After sequencing, phylogenetic tree analysis and protein secondary structure prediction were made. Results The results of quantitative analysis of serum HCV RNA before treatment showed that the virus titer in continuous response group (mean titer of 4 50 × 10 4 copies ml) was significantly lower than that in relapsed group and non-responsive group (mean titer of 1 82 × 10 7 copies ml) . 24 cases of chronic hepatitis C patients before treatment of interferon serum HCV ISDR amino acid sequence and anti-interferon HCV J strains compared to 13 cases of wild type, 11 cases of intermediate type, no mutation. Three of the six complete responders were infected with the wild-type strain and the other three were infected with the intermediate-type strain. Five strains of HCV NS5A full-length germline phylogenetic tree showed that three different types of response strains belong to three groups in the phylogenetic relationship, non-responders and interferon-resistant HCV J strain similar to 1 group. The prediction of protein secondary structure showed that the NS5A protein of the above strains were similar in secondary structure, with a significant difference only in the range of 2 2 55 ~ 2 2 89, which partially overlap with the PKR binding domain. Conclusion HCVNS5A gene?