基于限制性片段长度多态性的新方法对丙型肝炎病毒进行分型

Typing of hepatitis C virus by a new method based on restriction fragment length polymorphism.

作者信息

Buoro S, Pizzighella S, Boschetto R, Pellizzari L, Cusan M, Bonaguro R, Mengoli C, Caudai C, Padula M, Egisto Valensin P, Palù G

机构信息

Institute of Microbiology, University of Padova, Italy.

出版信息

Intervirology. 1999;42(1):1-8. doi: 10.1159/000024953.

DOI:10.1159/000024953

PMID:10393497

Abstract

A new restriction fragment length polymorphism (RFLP) analysis has been developed for hepatitis C virus (HCV) typing in the viral 5' non-coding region and contiguous core region. These genomic sequences were chosen for the relative nucleotide homology among different genotypes and for the presence of polymorphic sites. By employing two endonucleases (AccI and MboI) and, in some instances, a third one (EcoRII), we can unambiguously and reproducibly distinguish between genotypes and subtypes 1a, 1b, 1c, 2a, 2c, 2b, 3a, 3b, 4a, 5a and 6a. The method was applied for diagnosing two Italian groups of HCV-infected individuals reflecting a randomly collected population and a group of intravenous drug users. The accuracy of this method has been validated by comparison with INNOLiPA and by sequencing. Our approach represents an improvement over previous RFLP methods, since typing is accurate and simpler.

摘要

已开发出一种新的限制性片段长度多态性（RFLP）分析方法，用于对丙型肝炎病毒（HCV）的5'非编码区和相邻核心区进行基因分型。选择这些基因组序列是因为不同基因型之间存在相对核苷酸同源性以及存在多态性位点。通过使用两种核酸内切酶（AccI和MboI），在某些情况下还使用第三种核酸内切酶（EcoRII），我们可以明确且可重复地区分1a、1b、1c、2a、2c、2b、3a、3b、4a、5a和6a基因型及亚型。该方法应用于诊断两组意大利丙型肝炎病毒感染个体，一组为随机收集的人群，另一组为静脉注射吸毒者。通过与INNOLiPA比较和测序，验证了该方法的准确性。我们的方法是对先前RFLP方法的改进，因为基因分型准确且更简单。