2009年肠道病毒71型广州分离株的全基因组序列分析

The genome sequencing and analysis of an Enterovirus 71 strain isolated from Guangzhou in 2009

目的分析2009年肠道病毒71型广州分离株GZHY09的全基因组序列,并与GenBank中的序列进行分析比对。结论从GenBank上选不同地区的EV71全基因组序列,设计相互重叠的覆盖病毒整个基因组序列的12对引物,采用RT-PCR扩增出12个基因片段,通过测序获得全基因组序列,并参考国内外各EV71基因型的分离毒株,用MEGA4软件进行进化树分析,用DNAStar软件中的MegAlign进行同源性分析。结果 12个重叠基因片段序列拼接获得EV71全基因组序列,共7404个核苷酸,同源性分析结果表明在VP1区,GZHY09株与中国大陆的Anhui08、Zhejiang08、Shenzhen08的核苷酸序列同源性比较高,其中以Anhui08最高(97.7%)。结论 GZHY09属EV71病毒的C4亚型,与近年我国大陆地区流行的EV71株在进化上属于同一基因型,与Anhui08、Zhejiang08、Shenzhen08具有高度的同源性。

Objective To analyze the genome of an Enterovirus 71 isolate （strain GZHY09） obtained from Guangzhou in 2009 and to compare the sequences deposited in GenBank.The sequences were compared with the complete EV71 genome from the GenBank. Sequence analysis and gene identification were performed using bioinformatics software. Methods According to the genome information from GenBank, 12 pairs of primer were designed to cover the whole genome. The genome sequences were determined by RT-PCR sequencing method, and the sequences were analyzed using bioinformatic software （MEGA4 and DNASta）. Results The complete genome of GZHY09 contained 7404 nucleotides and the nucleotide sequence of VP-l regions was similar to the Anhui08, Zhejiang08 and Shenzhen08. Anhui08 had the highest sequence homology （97.7%） with GZHY09. Conclusion GZHY09 should be classified as subtype C4.GZHY09 is also phylogenitically similar to other epidemic strains found in the China's Mainland.