2013年至2018年辽宁省柯萨奇病毒A10型分离株VP1区基因特征分析

Genetic Characteristics of VP1 Region of Coxsackievirus A10 Strains in Liaoning Province during 2013-2018

为了解辽宁省地区手足口病患者中分离到的柯萨奇病毒(Coxsackievirus)A10型VP1区基因特征,收集了2013年至2018年辽宁省14个市送检的手足口病患者非EV-A71和非CV-A16肠道病毒的阳性标本,通过细胞培养法分离肠道病毒,提取病毒RNA;通过RT-PCR法进行肠道病毒VP1区基因扩增和序列测定;利用BLAST对测序结果比对后确定病毒基因型;对鉴定为CV A10的分离株进行VP1区同源性分析,并构建系统进化树。2013年至2018年辽宁省共收到非EV-A71和非CV-A16其他肠道病毒的阳性标本9 431份,用人横纹肌瘤细胞(rhabdomyosarcoma cells,RD)分离出CV-A10 165株。CV-A10分离株间的VP1区基因核苷酸同源性为91.3%~100%,氨基酸同源性为93.9%~100%,和A、B、C、D各基因型之间的VP1区同源性比较,与C基因型代表株的同源性最高,核苷酸同源性为92%~100%,氨基酸同源性为93.9%~100%。系统进化树分析表明,辽宁省CV-A10分离株为C基因型。 CV-A10是引起辽宁省手足口病的重要病原体,CV-A10分离株与C基因型代表株处于同一分支上均属C基因型,有C1和C2两个进化分支共同流行,其中C2分支是优势流行株。

In order to understand the genetic characteristics of Coxsackievirus A10 VP1 region isolated from patients with hand-foot-mouth-disease(HMFD)in Liaoning Province,the positive samples of non-EV71 and non-CV-A16 enteroviruses from patients with HMFD delivered for testing from 14 cities in Liaoning Province from 2013 to 2018.The enterovirus was isolated by cell culture method,viral RNA was extracted,and the VP1 region of enterovirus was amplified by RT-PCR and sequenced.The viral genotype was determined by comparing the sequencing results with BLAST.The VP1 region homology analysis was performed on the isolate identified as CV-A10,and a phylogenetic tree was constructed.The results showed that a total of 9431 clinical specimens of non-EV71 and non-CV-A16 enteroviruses were collected from Liaoning Province in 2013-2018.165 strains of CV-A10 were isolated from human rhabdomyosarcoma cells(RD).The VP1 region of the CV A10 isolate has a nucleotide homology of 91.3%-100%,amino acid homology of 93.9%-100%,andVP1 between the representative strains of A,B,C,and D genotypes.Compared with the C-type representative strain,the homology of the region was the highest,the nucleotide homology was 92%-100%,and the amino acid homology was 93.9%-100%.Phylogenetic tree analysis indicated that the CV-A10 isolate of Liaoning Province was a C genotype.CV-A10 was an important pathogen that caused HMFD in Liaoning Province.The CV-A10 isolate and the C genotype representative strain were all C genotypes on the same branch,and there were two evolutionary branches of C1 and C2.The C2 branch was the dominant strain.