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目的扩增HCV种属信息区NS5B和高度保守区5-UTR基因序列,用系统进化树法深入分析HCV基因型和亚型,阐明广东地区丙型肝炎患者HCV基因型和基因亚型的特点。方法采集99例慢性丙型肝炎病例血清,应用RT-PCR技术分别扩增HCVNS5B区和5-UTR特定片段,测序后进行系统进化树分析,确定HCV基因型及亚型。结果99例患者标本均准确分型,其中NS5B区段分型74例,5-UTR区段分型99例。HCV基因型和亚型的分布:1a亚型1例、1b亚型59例、1型7例、2a亚型11例、3a亚型3例、3b亚型2例、3型3例、6a亚型13例;另有2例标本在两区段分型不同,5-UTR/NS5B区段分型分别为6a/1b和3a/1b。两区段共同分型74例,72例标本在两区段基因型一致,2例标本在两区段基因型不同,为HCV重组体。结论联合分析HCV不同区段基因序列,可提高HCV基因分型的准确度及灵敏性。广东地区慢性丙型肝炎患者HCV基因亚型分布以1b为主;其次为6a和2a;3型中的a、b亚型也占有一定比例。
Objective To amplify the HCV genomic information region (NS5B) and highly conserved region 5-UTR (UTR) gene sequences and to analyze the HCV genotypes and subtypes using phylogenetic tree analysis to clarify the characteristics of HCV genotypes and genotypes in hepatitis C patients in Guangdong. Methods A total of 99 serum samples from patients with chronic hepatitis C were collected. The specific fragments of HCV NS5B and 5-UTR were amplified by RT-PCR and sequenced. Phylogenetic tree analysis was performed to determine the HCV genotypes and subtypes. Results All the 99 cases were correctly classified. Among them, 74 cases were classified by NS5B and 99 cases by 5-UTR. Distribution of HCV genotypes and subtypes: 1 in subtype 1a, 59 in subtype 1b, 7 in type 1, 11 in subtype 2a, 3 in subtype 3a, 2 in 3b, 3 in 3, and 6a Subtype in 13 cases. In the other two cases, the typing was different in the two sections. The 5-UTR / NS5B segments were classified as 6a / 1b and 3a / 1b, respectively. The two sections were co-located in 74 cases, 72 cases of the same genotype in the two sections, two cases of genotypic differences in the two sections for the HCV recombinant. Conclusion The combined analysis of HCV genomic sequences in different regions can improve the accuracy and sensitivity of HCV genotyping. HCV genotypes in patients with chronic hepatitis C in Guangdong were predominantly 1b, followed by 6a and 2a, and a and b subtypes in type 3 also accounted for a certain proportion.