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为了解藏族人群中流行的乙型肝炎病毒(HBV)基因型及基因重组状况,收集藏族人群HBsAg阳性样本,提取HBV DNA并用巢氏PCR的方法扩增HBV全长序列,测序后用DNAstar软件拼接全基因序列,进一步利用MEGA6和Simplot软件进行序列同源性和系统进化分析。应用该方法获得12株藏族人群HBV全基因序列,分析结果显示12株样本为C/D重组型,其中9株样本的重组位点位于nt750,3株样本的重组位点位于nt 1526,以此可分为两种重组方式,分别命名为C/Da和C/Db;从总体序列同源性来分析12株样本均属于C基因型,且与现有的C1-C15亚型的核苷酸差异均大于4%,与C1亚型最为接近。从地理分布来看,C/Da来自西藏中部和北部的拉萨市、阿里和林芝地区,C/Db均来自西藏南部的山南地区,提示两种重组型C/Da和C/Db在西藏地区的地理分布具有一定规律。研究结果可为西藏地区特殊HBV基因重组、基因特征分析、病毒进化研究以及当地乙肝防控提供参考。
In order to understand the prevalence of hepatitis B virus (HBV) genotypes and gene recombination among Tibetan people, HBsAg positive samples were collected from Tibetan populations, HBV DNA was extracted and full-length HBV sequences were amplified by nested PCR. After DNA sequencing software was sequenced, The whole genome sequences were further analyzed by sequence homology and phylogenetic analysis using MEGA6 and Simplot software. The HBV genome sequence of 12 Tibetans was obtained by this method. The results showed that 12 samples were C / D recombinants, of which 9 were located at nt750 and 3 were at nt 1526, They were divided into two recombination methods named as C / Da and C / Db, respectively. All 12 isolates were classified as C genotypes from the overall sequence homology and were similar to the existing C1-C15 subtype nucleotides Differences were greater than 4%, and C1 subtype closest to. In terms of geographical distribution, C / Da came from Lhasa, Ali and Nyingchi areas in central and northern Tibet, and C / Db from Shannan area in southern Tibet, suggesting that the two recombinant C / Da and C / Db in Tibet Geographical distribution has a certain law. The results provide a reference for the special HBV gene recombination, gene characterization, virus evolution research and local prevention and control of hepatitis B in Tibet.