摘　要:

[目的]建立引物特异性HCVRNA基因分型的方法，并应用于临床诊断。[方法]采用型特异性引物建立单管逆转录聚合酶链反应（RT-PCR）和双管引物特异性套式聚合酶链反应（NEST-PCR）检测技术，1b和3a一管内扩增，1a、2a和2b一管内扩增，分别对147例HCV抗体阳性的标本，设计HCV的C区特异性引物，采用Trizol浓缩RNA进行双管巢式PCR分型。[结果]统计结果表明我国丙型肝炎流行主要以1b（Ⅱ）和2a（Ⅲ）2种基因型为主，但有部分1a、2b、3a的HCV感染者检出，其中1a为1．36％（2／147），1b为65．99％（97／147），2a为6．12％（9／147），3a为1．36％（2／147），1b／2a型混合占22．45％（33／147），1a／2b混合型为0．68％（1／147），未分出型或其它型者分别为2．04％（3／147）。[结论]本文建立的方法具有良好的特异性和敏感性，可同时区分5种HCV基因型，我国北方地区发现有1a和3a基因型感染病例。

Establishment and application of genotype assay of HCV by dNest - PCR based on type - specific primers

Abstract:

[ Objective] To establish a method to type HCV by dNest - PCR based on type - specific primers. [ Methods] One tube RT -PCR and two tubes of Nest -PCR by specific -primer base on HCV C area （1b and 3a in 1 tube, 1a, 2a and 2b in another） were used to detect in samples from 147 HCV antibody -positive cases. [ Results] The 1b and 2a were the main genotypes of HCV in China. There were only 2 patients with 1a and 2 with 3a. For the 147 patients, the detecting rate of 1a, 1b, 2a, 3a, 1b mixed with 2a, 1a mixed with 2b and other genotypes was 1.36%, 65.99%, 6.12%, 1.36%, 22.45%, 0.68% and 2.04%, respectively. [ Conclusion] dNest- PCR based on genotype specific primers has high sensitivity and specificity. The genotypes of 1a and 3a are found in patients with HCV infection in North China.

Key words:

HCV; HCV typing by PCR with type -specific primers; RT- PCR; Nest -PCR