深圳献血人群隐匿性乙型肝炎病毒全基因序列系统进化树分析

目的了解深圳献血人群隐匿性乙型肝炎B和C基因型及亚型的分布规律,研究不同毒株全基因序列进化关系。方法对30例HBsAg(-)/HBV DNA(+)标本进行基本核心启动子/前C区(BCP/PC,295 bp)、HBV全基因(3 162 bp)巢式PCR扩增,PCR产物克隆后测序。2者均为阳性的5份标本合成3 215 bp长的全基因序列,与GenBank中已发表的HBV A～H基因型23株的全序列进行系统进化树分析确定基因型,将所属基因型B和C亚型再作系统进化树分析,以确定基因亚型。结果获得5例全基因序列,4例为C型,1例为B型。分属于C2,C2,C2,C1和B2亚型。3例C2亚型与日本、马来西亚基因亚型的进化距离最近,C1株与马来西亚和泰国2例携带者的病毒株进化距离最近。B2株与印度尼西亚华裔基因亚型的病毒株进化距离最近。结论深圳献血人群隐匿性乙型肝炎感染病毒基因亚型有C2,C1,B2,以C2亚型为主。

Analysis of phylogenetic tree of occult hepatitis B virus infection in Shenzhen blood donors

Objective To investigate the distribution of occult hepatitis B virus genotype B and C in Shenzhen blood donors and to study the phylogenetic relationship between the genotype B and C,and subgenotype B and C.Methods A total of 30 HBsAg(-)/DNA(+)plasma samples were amplified for Basic core promoter/pre-core region(BCP/PC,295 bp) and whole genome(3 162 bp) by using nested-PCR,and the PCR products were cloned,purified and then sequenced.Both BCP and the whole genome were obtained and combined HBV complete genome(3 21 5bp).Several full length sequences of HBV genotypes A to H and subgenotypes were obtained from the GeneBank database as reference sequences,and 5 full-length HBV genome sequence were determined genotype and subgenetype by phylogenetic analysis.Results Four of 5 OBI complete genome was genotype C,the other 1 was genotype B,and the subgenotype were C2,C2,C2,C1,and B2,respectively.The evalution distance of the 3 Chinese subtype C2 strain was the most hemogenous to Japan and Malaysia strain,and the C1 and B2 strain was to Malaysia and Thailand carrier strain and asymptomatic Chinese in Indonesia.Conclusion The HBV subgenotype in Shenzhen population is genotype C2,C1and B2,and the predominant type is C2.