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

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HCV RNA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/LR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HCV RNA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。

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

Objective To establish a method of nested RT-PCR to test HCV core protein gene and genotype.Methods HCV core protein gene sequence was searched from the Hepatitis C Virus（HCV） Database and HCV gene premier was designed for nested RT-PCR reaction system and genotyping.A total of 60 anti-HCV positive serum,50 anti-HCV negative and alanine aminotransferase（ALT） higher than 100U/L serum from entry-exit people were tested.Sequencing the RT-PCR reactive products for HCV genotype.Results From 60 anti-HCV positive serum,42（70%） were HCV core protein gene positive,including 27 lb genotype,12 la genotype,1 for 6a,3a,and 2a genotype.For 50 anti-HCV negative and ALT higher than 100 U/L serum,1 was 3a genotype.Conclusion This study established a quick and accurate method to test HCV core protein gene and genotype.