CAJ | 학술논문

目的：对获得的3株肠道病毒71（EV71）型毒株进行全基因组序列测定，并对其进化特点及分型进行初步分析。方法：提取病毒RNA，反转录得到cDNA，PCR分段扩增覆盖病毒全长序列的6个重叠片段（不包括多聚腺苷酸尾）；用软件将3株EV71的各片段序列进行拼接、编辑和校正，随后进行氨基酸翻译及序列比较；用MEGA4.1软件构建系统进化树。结果：获得了3株EV71的全长序列：GDV103株基因组全长7404 nt，包括741 bp的5'端非编码区（UTR）、6582 bp的病毒基因组编码区（ORF）及81 bp的3'UTR；安徽株（Anhui2007）基因组全长7405 nt，包括742 bp的5'UTR、6582 bp的ORF及81 bp的3'UTR；VR1432株基因组全长7408 nt，包括743 bp的5'UTR、6582 bp的ORF及83 bp的3'UTR长。经同源性比对和进化树分析，证实GDV103和安徽株EV71属于C基因型的C4基因亚型，而VR1432株则属于C基因型的C2基因亚型。结论：获得了3株EV71的全长基因组序列，并进一步探讨了其型别，为下一步的干扰素保护实验奠定了基础。
목적：대획득적3주장도병독71（EV71）형독주진행전기인조서렬측정，병대기진화특점급분형진행초보분석。방법：제취병독RNA，반전록득도cDNA，PCR분단확증복개병독전장서렬적6개중첩편단（불포괄다취선감산미）；용연건장3주EV71적각편단서렬진행병접、편집화교정，수후진행안기산번역급서렬비교；용MEGA4.1연건구건계통진화수。결과：획득료3주EV71적전장서렬：GDV103주기인조전장7404 nt，포괄741 bp적5'단비편마구（UTR）、6582 bp적병독기인조편마구（ORF）급81 bp적3'UTR；안휘주（Anhui2007）기인조전장7405 nt，포괄742 bp적5'UTR、6582 bp적ORF급81 bp적3'UTR；VR1432주기인조전장7408 nt，포괄743 bp적5'UTR、6582 bp적ORF급83 bp적3'UTR장。경동원성비대화진화수분석，증실GDV103화안휘주EV71속우C기인형적C4기인아형，이VR1432주칙속우C기인형적C2기인아형。결론：획득료3주EV71적전장기인조서렬，병진일보탐토료기형별，위하일보적간우소보호실험전정료기출。
Objective: To analyze the complete genomes of three enterovirus 71(EV71) strains and their sub-gen-otype. Methods: Six overlapping fragments covering the respective whole viral genome(excluding the polyA tail) were amplified by RT-PCR. The six DNA fragment sequences were spliced, edited and revised followed by transla-tion and sequence comparison. The phylogenetic tree was built using the MEGA4.1 software. Results: The full ge-nomic length of GDV103(7404 bp) was comprised of 5'UTR(741 bp), the open reading frame(ORF) 6582 bp and 3'UTR 81 bp. The full genomic length of Anhui2007 strain(7405 bp) was comprised of 5'UTR(742 bp), the ORF(6582 bp) and 3'UTR(81 bp). The full genomic length of VR1432(7408 bp) was comprised of 5'UTR(743 bp), the ORF(6582 bp) and 3'UTR(83 bp). The phylogenetic analysis based on the complete sequences showed that GDV103 and Anhui strain were classified into C4 sub-genotype and VR1432 into C2 sub-genotype. Conclu-sion: Three complete genomes of EV71 strains were obtained and the genotype was explored, which will facilitate the next interferon protection experiments.