2010-2012年扬州市手足口病病原构成及柯萨奇A组16型病毒分离株VP1基因特征分析  

Pathogen spectrum of hand-foot-and-mouth disease and genetic characteristics of VP1 from coxsackievirus A16 in Yangzhou from 2010 to 2012

作　　者：黄瑶[1,2] 周乐 巢国祥 芮明珠[3] 毕佩佩 卜平[2] 

机构地区：[1]扬州市疾病预防控中心,江苏扬州225001 [2]扬州大学医学院,江苏扬州225001 [3]扬州大学生物技术学院,江苏扬州225009

出　　处：《现代预防医学》2014年第13期2318-2322,2329,共6页Modern Preventive Medicine

基　　金：扬州市科技计划项目"肠道病毒EV71;CoxA16型特异性抗原表达及ELISA诊断试剂盒研制"(YZ2010090)

摘　　要：目的分析2010-2012年扬州地区手足口病的病原构成,并对柯萨奇A组16型（CA16）的VP1基因进行基因特征分析,了解扬州地区CA16病毒基因组VP1基因变化趋势。方法对2010-2012年从扬州市各县市区医院采集的手足口病临床标本1 309例进行肠道病毒荧光PCR检测,挑选56株CA16阳性标本进行病毒分离培养鉴定,并对6株CA16病毒分离株进行VP1基因测序,测序后利用DNAstar7.0软件进行基因比对,构建系统发生树。结果1 309份标本中检测到372份EV71阳性,295份CA16阳性,157份其他肠道病毒阳性。56株CA16阳性标本接种RD细胞获得30株CA16阳性毒株,测定其中6株病毒的VP1核苷酸序列,其同源性为96.4%-99.9%,氨基酸同源性为99.0%-100.0%,同GenBank中77株CA16参比毒株序列进化分析显示其属于B1b基因群。结论2010-2012年扬州市HFMD的主要病原为CA16（35.80%）和EV71（45.15%）,2011年以来其他肠道病毒比例显著上升。本地区分离CA16毒株同源性较高,与近年多数中国大陆分离株形成进化树上亲缘关系较近的一群,未出现明显毒力变化。Objective To understand the pathogen spectrum of HFMD in Yangzhou, analyze Coxsackievirus A 16（CA16） type VP1gene nucleotide and amino acid sequence, and understand the genetic characteristics of CA16 in Yangzhou. Methods 1 309 confirmed cases of HFMD specimens were collected from the hospitals in Yangzhou. All of the specimens were identified by real- time fluorescent PCR with EV71 and CA16-type-specific primers. CA16 isolations were conducted with rhabdomyosarcoma（RD） cell line on 56 throat swabs having CA 16 positive realtime-PCR results.Full length VP1 genes from these six CA16 strains were amplified by RT-PCR and sequenced. The sequences were compared with VP1 genes from CA16 in GenBank database. Sequence analysis and type identification, and the phylogenetic tree construction were performed by DNAstar7.0. Results Of totally 1 309 HFMD cases, 372 were EV71 positive, 295 were CA16 positive, 157 were other Enteroviruses positive. 30 CA16 isolations were obtained and the VP1genes of 6 CA16 isolations were sequenced and analyzed. The nucleotide and amino acid homogeneity between of these6 Yangzhou strains ranged from 96.4% to 99.9% and 99.0% to 100%. The sequences compared with VP1 genes from 77 strains of CA16 in GenBank database showed that the CoxA16 strains prevalent in Yangzhou from 2010 to 2012 belonged to subgenotype B1b. Conclusion The prevalent pathogens of HFMD in Yangzhou are CA16（35.80%） and EV71（45.15%）. The proportion of other enteroviruses increases significantly since 2011, and the isolates in this study have close association in genetic relationship, and are closely related with some mainland Chinese isolates in recent years.There is still no evidence that this separation in Yangzhou 6CA16 strains have enhanced virulence.

关 键 词：手足口病 柯萨奇病毒A组16型(CA16) VP1基因 

分 类 号：R183[医药卫生—流行病学]