出入境人员丙型肝炎病毒核心蛋白基因巢式RT-PCR检测及分型研究

    古莉冰1，朱玉兰1，徐媛1，孙杰1，王佃鹏2，董瑞玲1

    1.深圳国际旅行卫生保健中心，广东 深圳 518033;2.深圳市职业病防治院

    摘要：目的建立一种检测丙型肝炎病毒（HCV）核心蛋白基因的巢式RT-PCR方法，以及HCV核心蛋白基因区测序分型方法。方法从国际HCV数据库查找HCV核心蛋白基因序列，设计HCVRNA巢式RT-PCR引物及测序引物，并建立巢式RT-PCR反应体系。采用出入境人员抗-HCV阳性血清60份，及50份标本为HCV抗体检测阴性，且谷丙转氨酶（ALT）＞100U/L的血清进行巢式RT-PCR检测，阳性结果使用本研究建立的测序引物进行基因测序以确定HCV基因型。结果60份抗-HCV阳性血清中，42份（70.0%）核心蛋白基因RT-PCR扩增阳性，其中1b型27例、1a型12例、6a型1例、3a型1例、2a型1例。50份ALT＞100U/L且抗-HCV阴性标本中，1份HCV核心蛋白基因RT-PCR扩增阳性,测序鉴定为3a型。结论 本研究建立了一种HCVRNA核心蛋白基因的巢式RT-PCR检测和分型方法，为出入境人员HCV的诊断和监测提供重要依据。

    关键词：丙型肝炎病毒；核心蛋白基因；巢式RT-PCR；基因分型

    中图分类号：R512.6　文献标识码：B

    A nested RT-PCR method of HCV RNA coreprotein gene testing and genotyping for entry-exit people

    GU Li-bing*, ZHU Yu-lan, XU Yuan, SUN Jie,WANG Dian-peng, DONG Rui-ling

    *Shenzhen International Travel HealthcareCenter, Shenzhen, Guangdong 518033, China

    Abstract:   Objective  To establish a method of nested RT-PCR to test HCV core proteingene and genotype.  Methods   HCV core protein genesequence was searched from the Hepatitis C Virus (HCV) Database andHCV gene premier was designed for nested RT-PCR reaction system andgenotyping. A total of 60 anti-HCV positive serum，50 anti-HCVnegative and alanine aminotransferase (ALT) higher than 100U/Lserum from entry-exit people were tested. Sequencing the RT-PCRreactive products for HCV genotype.  Results   From60 anti-HCV positive serum，42(70%) were HCV core protein genepositive, including 27 1b genotype, 12 1a genotype, 1 for 6a, 3a,and 2a genotype. For 50 anti-HCV negative and ALT higher than 100U/L serum, 1 was 3a genotype.  Conclusion   Thisstudy established a quick and accurate method to test HCV coreprotein gene and genotype.

    Key words:   Hepatitis C Virus;Core protein gene; Nested RT-PCR; Genotyping

    《中国国境卫生检疫杂志》