中华肝脏病杂志
中華肝髒病雜誌
중화간장병잡지
CHINESE JOURNAL OF HEPATOLOGY
2012年
10期
766-768
,共3页
肝炎病毒,戊型%开放读码框3%核苷酸同源性%系统进化分析
肝炎病毒,戊型%開放讀碼框3%覈苷痠同源性%繫統進化分析
간염병독,무형%개방독마광3%핵감산동원성%계통진화분석
Hepatitis E virus%Open reading frame 3%Sequence homology,nucleotide%Phylogenetic analysis
目的 对武汉地区戊型肝炎病毒(HEV)开放读码框(ORF)3基因序列进行分析,并确定病毒基因型. 方法 收集103份抗-HEV IgM阳性血清,采用逆转录-巢式聚合酶链反应扩增HEV RNA两个基因片段(5020 ～ 5392nt和5347 ～ 5956 nt,EF570133)；对PCR产物进行测序,并用ContigExpress将测序结果拼接(含有ORF3基因),对ORF3基因序列进行分析.结果 103份抗-HEV IgM阳性血清中HEV RNA两个基因片段均扩增出来的样本为18份,18株HEV ORF3基因全长均为345 bp,编码114个氨基酸.各毒株间的核苷酸同源性为92.5％ ～ 99.4％,与基因Ⅰ型、Ⅱ型、Ⅲ型、Ⅳ型HEV的核苷酸同源性分别为83.5％ ～ 86.7％、83.2％ ～ 85.2％、84.6％ ～ 87.2％、92.0％ ～ 96.5％;系统进化分析表明该18株HEV均为基因Ⅳ型. 结论 武汉地区HEV主要为基因Ⅳ型,ORF3基因序列可用于同源进化分析.
目的 對武漢地區戊型肝炎病毒(HEV)開放讀碼框(ORF)3基因序列進行分析,併確定病毒基因型. 方法 收集103份抗-HEV IgM暘性血清,採用逆轉錄-巢式聚閤酶鏈反應擴增HEV RNA兩箇基因片段(5020 ～ 5392nt和5347 ～ 5956 nt,EF570133)；對PCR產物進行測序,併用ContigExpress將測序結果拼接(含有ORF3基因),對ORF3基因序列進行分析.結果 103份抗-HEV IgM暘性血清中HEV RNA兩箇基因片段均擴增齣來的樣本為18份,18株HEV ORF3基因全長均為345 bp,編碼114箇氨基痠.各毒株間的覈苷痠同源性為92.5％ ～ 99.4％,與基因Ⅰ型、Ⅱ型、Ⅲ型、Ⅳ型HEV的覈苷痠同源性分彆為83.5％ ～ 86.7％、83.2％ ～ 85.2％、84.6％ ～ 87.2％、92.0％ ～ 96.5％;繫統進化分析錶明該18株HEV均為基因Ⅳ型. 結論 武漢地區HEV主要為基因Ⅳ型,ORF3基因序列可用于同源進化分析.
목적 대무한지구무형간염병독(HEV)개방독마광(ORF)3기인서렬진행분석,병학정병독기인형. 방법 수집103빈항-HEV IgM양성혈청,채용역전록-소식취합매련반응확증HEV RNA량개기인편단(5020 ～ 5392nt화5347 ～ 5956 nt,EF570133)；대PCR산물진행측서,병용ContigExpress장측서결과병접(함유ORF3기인),대ORF3기인서렬진행분석.결과 103빈항-HEV IgM양성혈청중HEV RNA량개기인편단균확증출래적양본위18빈,18주HEV ORF3기인전장균위345 bp,편마114개안기산.각독주간적핵감산동원성위92.5％ ～ 99.4％,여기인Ⅰ형、Ⅱ형、Ⅲ형、Ⅳ형HEV적핵감산동원성분별위83.5％ ～ 86.7％、83.2％ ～ 85.2％、84.6％ ～ 87.2％、92.0％ ～ 96.5％;계통진화분석표명해18주HEV균위기인Ⅳ형. 결론 무한지구HEV주요위기인Ⅳ형,ORF3기인서렬가용우동원진화분석.
Objective To determine the distribution ofgenotype Ⅳ among hepatitis E virus (HEV)infections in Wuhan by sequencing the open reading frame (ORF) 3 gene of HEV clinical isolates.Methods Serum samples were collected from 103 individuals who tested positive for the anti-HEV IgM antibody,and total HEV RNA was extracted for targeted gene sequencing analysis.Reverse transcription-nested polymerase chain reaction (PCR) was used to amplify two fragments of the ORF3 gene (5020 ～ 5392 nt and 5347 ～ 5956nt,EF570133).The two PCR products were sequenced and the sequences were stitched with the ContigExpress program and used to determine the HEV genotype.Results Both ORF3 gene fragments were amplified in 18 out of the 103 anti-HEV IgM-positive serum samples.These 18 HEV isolates shared 92.5％ ～ 99.4％ identity with each other at the nucleotide level.Nucleotide sequence homology analysis of the HEV genotypes Ⅰ,Ⅱ,Ⅲ,and Ⅳ indicated the highest homology was with genotype Ⅳ; specifically,homology with genotype Ⅰ was 83.5％ ～ 86.7％,with genotype Ⅱ was 83.2％ ～ 85.2％,with genotype Ⅲ was 84.6％ ～ 87.2％,and with genotype Ⅳ was 92.0％～ 96.5％.Conclusion Targeted sequencing of the HEV ORF3 gene facilitated genotyping of clinical isolates.Using this method,it was determined that nearly 20％ of HEV clinical isolates from Wuhan belong to genotype Ⅳ.
