3株肠道病毒71型毒株的全基因组序列分析

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eDNA，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的备片段序列进行拼接、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741bp的5’端非编码区（UTR）、6582bp的病毒基因组编码区（ORF）及81bp的3’UTR；安徽株（Anhui2007）基因组全长7405nt，包括742bp的5＇UTR、6582bp的ORF及81bp的3＇UTR；VR1432株基因组全长7408nt，包括743bp的5’UTR、6582bp的ORF及83bp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宴，哈重定了基础．

Complete Genomic Analysis of Three Enterovirus 71 Strains

Objective： To analyze the complete genomes of three enterovirus 71（EV71） strains and their sub-genotype. 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 translation and sequence comparison. The phylogenetic tree was built using the MEGA4.1 software. Results： The full genomic length of GDVI03（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 GDVI03 and Anhui strain were classified into C4 sub-genotype and VR1432 into C2 sub-genotype. Conclusion： Three complete genomes of EV71 strains were obtained and the genotype was explored, which will facilitate the next interferon protection experiments.