从1例急性弛缓性麻痹患儿分离到的埃可病毒33型全基因组序列分析  

Whole-genome sequence analysis of echovirus type 33 isolated from a child with acute flaccid paralysis

作　　者：袁芳[1] 周娟[1] 曹慜 张薇 马学旻[1] 马江涛[1] 魏新峰[2] 马学平[1] 王青利 YUAN Fang;ZHOU Juan;CAO Min;ZHANG Wei;MA Xuemin;MA Jiangtao;WEI Xinfeng;MA Xueping;WANG Qingli(Ningria Hui Autonomous Region Center for Disease Prevention and Control,Yinchuan,Ningria 750004,China;不详)

机构地区：[1]宁夏回族自治区疾病预防控制中心,宁夏银川750004 [2]宁夏回族自治区人民医院,宁夏银川750002 [3]西安安琪儿妇产医院,陕西西安710077

出　　处：《中国病毒病杂志》2023年第6期436-442,共7页Chinese Journal of Viral Diseases

基　　金：国家自然科学基金(81960607);宁夏自然科学基金(2021AAC03415,2022AAC03708)。

摘　　要：目的了解2015年宁夏地区埃可病毒33型(echovirus type 33,E33)分离株基因特征,为今后的科学防控提供参考。方法将2015年急性弛缓性麻痹(acute flaccid paralysis,AFP)监测系统中收集到的病例粪便标本和密切接触者粪便标本按照监测方案要求进行处理后,接种于人恶性胚胎横纹肌瘤(human rhabdomyosarcoma,RD)细胞,分离获得病毒后,提取病毒RNA,通过扩增肠道病毒完整VP1区序列鉴定型别,选取2株采用Illumina全基因测序方法进行全基因组序列测定。用MegaX、DNAstar进行遗传进化分析,利用RDP4和SimPlot3.5.1进行重组分析。结果分离的4株毒株均扩增到完整VP1区序列,鉴定为E33病毒,将测序获得的2株全基因组序列与GenBank中下载的E33原型代表株Toluca-3的全基因组序列比较,核苷酸相似性为79.9%,氨基酸同源性为95.7%。基于E33 VP1区843 bp核苷酸序列构建系统进化树,宁夏地区分离到的E33分离株属于C2亚型,与湖南、江苏、新疆等地流行的E33属于同一亚型,氨基酸序列同源性和核苷酸序列相似性较高。基于全基因组序列和P1、P2、P3区序列分别构建系统进化树,结果显示宁夏地区流行的E33在全基因组进化树上与新疆地区流行的EV-B85聚类,未与E33原型株聚类。宁夏地区流行的E33在P1区基因进化树上与E33原型株聚类,在P2、P3区基因进化树与EV-B85聚类。用RDP4和Sim Plot3.5.1软件进行重组分析后证实宁夏地区流行的E33为重组株,在P1区与E33原型株重组,在P3区与EV-B85重组。结论宁夏地区流行的E33为E33原型株和EV-B85重组形成的新的重组病毒,有关该重组株的致病性有待进一步的研究。Objective To understand the genetic characteristics of a strain of human echovirus type 33(E33)isolated in Ningxia Hui autonomous region in 2015,so as to provide reference for future scientific prevention and control.-Methods The fecal samples of a case and contacts collected in the acute flaccid paralysis(AFP)monitoring system in 2015 were processed following the method required by the monitoring protocol,inoculated on RD cells to obtain virus.The viral RNA was extracted and identified,by amplifying the sequence of complete VPl sequence of enterovirus.Two strains were selected for whole-genome sequencing using Ilumina.Genetic evolution analysis was performed with MegaX and DNAstar,and recombination analysis with RDP4 and SimPlot3.5.1R.esults Complete VPl region sequence was successfully amplified in all the four isolated strains,which were identified as E33 virus.Comparison of the whole genome sequences of the two strains obtained by sequencing with the whole genome sequences of the E33 prototype strain Toluca-3 downloaded from GenBank.showed a nucleotide homology of 79.9% and an amino acid homology of 95.7%.According to the polygenetic tree constructed based on the 843 bp nucleotide sequence of the E33 VP1 region,the E33 isolates in Ningxia belonged to the C2 lineage,which was the same lineage as E33 circulating in Hunan,Jiangsu and Xinjiang,and had high amino acid homology and nucleotide homology.The polygenetic tree constructed based on the whole-genome sequence showed that the E33 in Ningxia clustered with the EV-B85 circulating in Xinjiang,but not with the E33 prototype strain.It clustered with the E33 prototype strain in the polygenetic tree based on Pl region,and with the EV-B85 in the polygenetic trees based on P2 and P3 regions respectively.Recombination analysis with RDP4 and SimPlot 3.5.1 confirmed that the isolates in Ningxia were recombinant strains,which recombined with the E33 prototype strain in the P1 region,and with the EV-B85 strain in the P3 region.Conclusion The E33 circulating in Ningxia is

关 键 词：急性弛缓性麻痹 肠道病毒 全基因组 遗传进化分析 重组分析 

分 类 号：R512.5[医药卫生—内科学]