猪源戊型肝炎病毒上海株全基因组序列分析  

Analysis of the complete genome of a swine strain of hepatitis E virus isolated from Shanghai

作　　者：曾容愚[1,2] 刘佩红[1] 周锦萍[1] 张维谊[1] 张苏华[1] 陆承平[2] 

机构地区：[1]上海市动物疫病预防控制中心,上海201103 [2]南京农业大学,南京200095

出　　处：《中国人兽共患病学报》2009年第2期165-168,共4页Chinese Journal of Zoonoses

摘　　要：目的分析猪源戊型肝炎病毒(HEV)上海株swChinash与已知其它HEV基因型的差异和人源HEV的关系。方法设计22对PCR引物,分片段进行扩增,两末端采用快速扩增方法(RACE)扩增,扩增产物克隆到pMD18-T载体并测序,用DNASTAR软件与53株人源和猪源HEV的基因组进行同源性分析,用Clustalx和Mega3.0软件绘制基因进化树。结果swChinash全序列除去poly(A)尾,由7235个核苷酸组成,ORF1～3分别编码1707,660,122个氨基酸。全基因序列与Ⅳ型的核苷酸同源性为83.4%～84.7%,与其他三型为73.3%～76.3%,其中与中国人源长春株最高,为84.7%。结论基于ORF1～3和全长基因组的进化树,猪源HEV上海株swChinash属于基因Ⅳ型,与其他Ⅳ型HEV亲缘关系较远,单独在一个进化分支上。To analyze the full length sequence of swine strain of hepatitis E virus（HEV） swChinash, 11 overlapping fragments of swChinash genomes were amplified with reverse transcription nested polymerase chain reaction（RT-nPCR） and the 5＇and 3‘ends were amplified with RACE. The PCR products were cloned into Pmd18-T vector and sequenced. The result showed that its genome consisted of 7 235nt excluding the ploy（ A） tail, and contained three ORFs（ORF1--3）that encoded proteins of 1707,660,122aa. sequence analysis revealed that the overall nucleotide sequence identity of swChinash was 73.3% -76.3% and 83.4%-84.7% with HEV genotype Ⅰ -Ⅲ and Ⅳ ,respectively. It shared 84.7% nucletotide identity with China 8 human strain. Necletotide identity analysis based on three ORFs and the full length sequence yielded similar results： it belongs to genotype sⅣ. However, phylogenetic analyses suggests that it represents a distinct variant among the genotype Ⅳ isolates.

关 键 词：戊型肝炎病毒 RT-nPCR 序列分析 基因型 

分 类 号：R373[医药卫生—病原生物学]