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目的了解2010年长沙市手足口病(hand-foot-mouth disease,HFMD)重症和普通病例肠道病毒71型(human Enterovirus 71,EV71)VP1区基因特征。方法收集12例HFMD重症和普通病例的咽拭子或肛拭子标本进行VP1区基因RT-PCR扩增和核苷酸序列测定,测序结果利用sequin软件提交至GenBank,Lasergene和Mega5软件对测序结果进行氨基酸比对分析和构建进化树。结果 12株EV71 VP1区基因序列在进化树上与C基因型中的C4a亚型代表株属同一分支,在核苷酸同源性分析中,12株EV71与C4a亚型代表株(3254-TAI-98)的同源性达到92.6%~93.3%,高于与A、B、C1、C2、C3、CVA16型或亚型代表株的同源性(分别为79.9%~80.7%、83.1%~84.0%、87.9%~88.7%、89.1%~89.8%8、7.8%~88.2%和57.3%~58.4%);12株EV71病毒VP1之间的核苷酸有高度的同源性:同源性为98.1%~100.0%,高于与C4亚型代表株的同源性(92.6%~93.3%和91.8%~92.7%)。2010年分离自长沙的重症和普通病例与2008年分离自北京、深圳和阜阳的EV71病毒VP1基因序列之间的氨基酸变异位点少。结论 2010年长沙市流行的EV71型病毒属于C基因型中的C4亚型;12例HFMD重症和普通病例的EV71 VP1区基因序列差异较小;不同时间、不同地区和不同病例来源的EV71 VP1区氨基酸变异位点无关联。
Objective To understand the characteristics of VP1 gene in severe and common cases of hand-foot-mouth disease (HFMD) in Changsha in 2010. Methods The throat swabs or anus swab samples from 12 severe and common HFMD cases were collected for RT-PCR amplification and nucleotide sequencing of VP1 gene. The sequencing results were submitted to GenBank, Lasergene and Mega5 software using sequin software for sequencing results Amino acid alignment analysis and construction of phylogenetic trees. Results The sequences of the 12 EV71 VP1 genes shared the same branch with the C4a subtype of C genotype in the phylogenetic tree. Among the nucleotide homology analysis, 12 EV71 and C4a subtypes (3254-TAI -98) were 92.6% -93.3% higher than that of the representative strains of A, B, C1, C2, C3 and CVA16 or subtype (79.9% -80.7%, 83.1% 84.0%, 87.9% -88.7%, 89.1% -89.8% 8,7.8% -88.2% and 57.3% -58.4%, respectively). There was a high degree of homology between nucleotides of VP1 of 12 EV71 viruses: homology Was 98.1% -100.0%, higher than that of the C4 subtype (92.6% -93.3% and 91.8% -92.7%). In 2010, severe and common cases isolated from Changsha and the 2008 EV71 virus isolated from Beijing, Shenzhen and Fuyang virus VP1 gene sequence between the amino acid mutation sites less. CONCLUSIONS: The EV71 virus in Changsha City belonged to the C4 subtypes of C genotype in 2010; EV71 VP1 gene sequences of 12 severe and common cases of HFMD were relatively small. EV71 VP1 region in different regions, different cases and different cases Amino acid variation sites are not related.