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+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′(BsrB Ⅰ、HaeⅡ、Hinf Ⅰ)复合内切酶消化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HaeⅢ内切酶鉴别2a、2b、3b、4a及6a。电泳检测片段大小。结果 (1)1a、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′-NCRABC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。
Objective To further understand whether HCV 3b and genotypes 1a, 2b and 6a are present in China, a genomic library of different genotypes of 5’-NCR in 5’-untranslated region of HCV was constructed. METHODS: The genotyping method was performed according to ABC program. A digested 5’-NCR cDNA with BHH ’(BsrB Ⅰ, HaeⅡ, Hinf Ⅰ) endonuclease and divided the genotypes into 5 groups: 1a, 1b, 6a, 2a , 2b, 3a, 3b, 4a. B application of BstU Ⅰ endonuclease identification 1a, 1b. C Use Hae III endonucleases to identify 2a, 2b, 3b, 4a and 6a. Electrophoresis detection fragment size. Results (1) ABC typing of 8 genotypes 1a, 1b, 2a, 2b, 3a, 3b, 4a, 6a showed that these 8 genotypes had good typing results. (2) ABC typing of 93 HCV RNA-positive patients showed that the infection rate of type 1b was 66.67%, type 2a was 18.28%, type 1b / 2b, type 3b and type 2b were 3.23%, type 2a / 2b and type 1b / 2a type each 2.15%, 1a type 1.08%. Conclusion The results show that using HCV 5’-NCRABC typing technology not only guarantees the sensitivity of HCV RNA detection, but also completes the identification of 8 genotypes in 1a ~ 6a.