丙型肝炎病毒非编码区ABC程序酶切分型研究

目的为进一步了解中国是否存在HCV 3b基因及1a、2b和6a基因型感染，建立HCV 5′端非编码区(5′ NCR)不同基因型的基因库。方法分型方法按ABC程序进行，A应用BHH′(BsrBⅠ、HaeⅡ、HinfⅠ)复介内切酶消化5′NCR cDNA，可将不同基因型划分为5组：1a、1b，6a，2a、2b，3a，3b、4a。B应用BstU Ⅰ内切酶鉴别1a、1b。C应用Hae Ⅲ内切酶鉴别2a、2b、3b、4a及6a。电泳检测片段大小。结果(1)la、1b、2a、2b、3a、3b、4a、6a 8种基因型参比品的ABC分型结果表明，该8种基因型获得良好的分型效果。(2)93份HCV RNA阳性患者ABC分型结果表明，1b型感染率占66．67％，2a型18．28％，1b／2b型、3b型及2b型均为3．23％，2a／2b型和1b／2a型各为2．15％，1a型1．08％。结论结果表明应用HCV 5′-NCR ABC分型技术既保证了HCV RNA检测的灵敏度，又能完成1a-6a型中的8种基因型的鉴别。

Hepatitis c virus genotype research by ABC programs of 5'-NCR restriction endocuclease digestion

OBJECTIVE: In order to fully understand hepatitis c virus (HCV) genotype 3b, 1a, 2b and 6a infection in China, We built HCV 5'-noncoding region (5'-NCR) of different genotypes and subtypes. METHODS: The classification HCV into variable genotypes (subtypes) was carried on by programs A, B and C A. Using a combination of three restriction endonuclease BHH' (BsrB I, Hae II, Hinf I) digestions at the same time. The distinct genotypes were classified into 5 groups: genotype 1 (1a, 1b), 6a, 2 (2a, 2b), genotype 3 (3a, 3b), genotype4 (4a). B. With regard to genotype 1, we could distinguish subtype 1a from 1b using BstU I digestion. C. Using restriction endonuclease Hae III, genotype 2a, 2b, 3b, 4a, 6a are differentiated respectively. RESULTS: (1) HCV genotype 1a, 1b, 2a, 2b, 3a, 3b, 4a, 6a are fully discriminated by comparison with the genotypes regular samples. (2) Of the 93 patients, HCV genotype distribution in China was 66.67% for 1b, 18.28% for 2a, 3.23% for 1b/2b, 3b, 2b respectively. 2.15% for 2a/2b, 1b/2a respectively. 1.08% for 1a. CONCLUSION: This research indicated that adoption of HCV 5'-NCR A B C restriction endonuclease digestions techniques, might be sensitive and efficient to detect HCV and discriminate HCV genotype (subtypes) 1a to 6a.