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目的通过扩增VP1节段的核苷酸序列,对2008年珠海暴发流行的EV 71进行种系进化分析。方法选取10例手足口病暴发流行中的EV71阳性病例标本用一对特异引物进行VP1全基因核苷酸序列扩增,由生物技术公司在ABI3730型自动测序仪上测序,序列结果用Claxter、DNA Star、Mega软件进行分析。结果本文的10株病毒与C基因型代表株比较接近,核苷酸同源性在88.2%～99.2%之间,氨基酸同源性在98.7%～99.7%之间;尤其与C4基因型最为相近,与C4基因型相比较核苷酸同源性在97.6%～99.2%之间,氨基酸同源性在99.3%～99.7%之间;而与A、B基因型代表株比较差异较大,核苷酸同源性只在81.8%～84.4%之间,氨基酸同源性在94.6%～98.0%之间;说明本文的10株病毒皆属于EV71病毒C基因型、C4亚型。结论珠海和新会的病毒株可能与阜阳和浙江分离的EV71病毒有相同的起源,均属于C4亚型,并且C4亚型病毒在中国大陆可能有较广泛的分布和传播,因此加强各地流行的EV71病毒基因特征分析,进而探索EV71病毒的可能进化途径,对于肠道病毒的基础研究和防范EV71在中国的大规模暴发与流行具有重要意义。
OBJECTIVE: To amplify the nucleotide sequence of VP1 segment, phylogenetic analysis was performed on the EV 71 epidemic in Zhuhai in 2008. Methods A total of 10 EV71-positive cases of HFMD outbreaks were selected to amplify the full-length VP1 gene by using a pair of specific primers. Sequencing was performed on biotype ABI 3730 automatic sequencer using Claxter, DNA Star, Mega software for analysis. Results The 10 isolates of this study were relatively close to the C genotype, with nucleotide homology of 88.2% -99.2% and amino acid identities of 98.7% -99.7%, especially those of C4 genotype Compared with the C4 genotype, the nucleotide homology was between 97.6% -99.2% and the amino acid homology was between 99.3% -99.7%, while it was significantly different from that of the representative genotypes A and B The homology of the nucleotide sequences was only between 81.8% and 84.4%, and the amino acid homology was between 94.6% and 98.0%. It indicated that all the 10 viruses in this study belong to genotype C and C4 of EV71 virus. Conclusion The strains of Zhuhai and Xinhui may share the same origin with the EV71 viruses isolated in Fuyang and Zhejiang and all belong to the C4 subtype and the C4 subtype viruses may have a broader distribution and spread in mainland China, EV71 virus gene characteristics, and then explore the EV71 virus may evolve ways for the basic research of enterovirus and EV71 in China to prevent large-scale outbreaks and epidemic of great significance.