淮南市手足口病病毒分离株VP1基因分子特征分析

发布时间：2018-06-29 18:29

本文选题：手足口病 + EV　；参考：《中国病原生物学杂志》2017年12期

【摘要】：目的了解淮南市手足口病病毒分离株VP1基因遗传特征及进化关系。方法用人横纹肌肉瘤(rhabdomyosarcoma,RD)细胞对2014-2016年荧光PCR检测为EV71型、CA16型及CA10型阳性标本进行病毒分离,经2代接种出现细胞病变(CPE)后取培养物上清,提取病毒RNA,RT-PCR法扩增不同基因型VP1基因并测序,采用Bioedit5.0.6、DNAstar7.1和Mega 6.06生物软件对测序结果进行比对及进化分析。结果 11株EV71型分离株VP1编码区的核苷酸片段长度为891nt,同源性为93.5%-100.0%,氨基酸同源性为98.3%-100.0%;7株CVA16型分离株VP1编码区的核苷酸片段长度为891nt,同源性为95.1%-97.0%,氨基酸同源性为98.7%-100.0%;3株CVA10型分离株VP1编码区的核苷酸片段长度为894nt,同源性为96.5%-100.0%,氨基酸同源性则为98.7%-100.0%。系统进化分析EV71型淮南株与安徽阜阳、山东、河南、北京、云南分离毒株亲缘性较近,处于进化分支C4a,核苷酸同源性为93.5%-100.0%,氨基酸同源性为98.7%-100.0%;CVA16型淮南株为B1b亚型,与山东株、河南株、深圳株、南京株亲缘关系较近,核苷酸序列同源性为95.1%-99.1%,氨基酸序列同源性98.7%-100.0%;CVA10淮南株处于D分支内,与云南、河南、河北株亲缘关系较近,核苷酸序列同源性为96.5%-100.0%,氨基酸序列同源性98.3%-100.0%。结论淮南市2014-2016年流行的EV71为C4a亚型,CVA16为B1b亚型,CVA10处于D聚类分支,与国内其他地区流行的手足口病毒亚型一致。
[Abstract]:Objective to investigate the genetic characteristics and evolutionary relationship of VP1 gene of HFMD virus isolate in Huainan City. Methods Human rhabdomyosarcomac Rd cells were isolated from positive specimens of EV71 type CA16 and CA10 detected by fluorescent PCR from 2014-2016. The supernatant of culture was obtained after inoculation with cytopathic acid (CPE) for 2 passages. The VP1 gene of different genotypes was amplified by RT-PCR and sequenced. Bioedit5.0.6 DNA star7.1 and Mega 6.06 were used to compare and analyze the sequencing results. Results the nucleotide fragment length of VP1 coding region in 11 EV71 isolates was 891nt.The homology was 93.5-100.0.The amino acid homology was 98.3-100.00.The nucleotide length of VP1 coding region of 7 CVA16 isolates was 891nt, the homology was 95.1 -97.0, and the amino acid homology was 98.7- 100.0.The amino acid homology was 98.7- 100.020. The nucleotide fragment length of VP1 coding region of CVA10 isolate was 894nt.The homology was 96.5- 100.0.The amino acid homology was 98.7- 100.0. Phylogenetic analysis showed that Huainan strain EV71 was closely related to the isolates isolated from Fuyang, Shandong, Henan, Beijing and Yunnan, and was in the evolutionary branch of C4a. The nucleotide homology was 93.5-100.0.The amino acid homology was 98.7-100.0B1b subtype of Huainan strain of CVA16, which was subtype of B1b with Shandong strain. The homology of nucleotide sequence of Henan strain, Shenzhen strain and Nanjing strain was 95.1 -99.1, the amino acid sequence homology was 98.7-100.00.The Huainan strain of CVA10 was in D branch, and had close relationship with Yunnan and Hebei strains. The homology of nucleotide sequence was 96.5- 100.0.The homology of amino acid sequence was 98.3- 100.0km. Conclusion the EV71 epidemic in Huainan City from 2014-2016 is C4a subtype, CVA16 is B1b subtype, CVA10 is in D cluster branch, which is consistent with the HFMD virus subtype prevalent in other parts of China.
【作者单位】：淮南市疾病预防控制中心;安徽省疾病预防控制中心;
【基金】：淮南市市级科技计划项目(No.1307)
【分类号】：R440;R725.1

【相似文献】