论文部分内容阅读
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 analyze the pathogenic constitution of HFMD in Yangzhou area from 2010 to 2012 and analyze the gene characteristics of VP1 gene of Coxsackie A group type 16 (CA16) to find out the change trend of VP1 gene in CA16 virus genome of Yangzhou area. Methods A total of 1 309 clinical samples of hand-foot-mouth disease collected from urban counties in Yangzhou City from 2010 to 2012 were detected by real-time PCR. Fifty-six CA16 positive samples were selected for virus isolation and culture. Six strains of CA16 virus The VP1 gene was sequenced and sequenced. DNAstar7.0 software was used for gene alignment to construct phylogenetic tree. Results A total of 372 positive samples of EV71, 295 positive samples of CA16 and 157 positive samples of other enteroviruses were detected in 309 samples. Fifty-six CA16 positive specimens were inoculated into RD cells to obtain 30 CA16 positive strains. The VP1 nucleotide sequences of 6 strains of viruses were detected. The homology was 96.4% -99.9% and the amino acid homology was 99.0% -100.0% The phylogenetic analysis of the 77 CA16 reference strains in GenBank revealed that they belonged to B1b gene group. Conclusion The main pathogens of HFMD in Yangzhou from 2010 to 2012 were CA16 (35.80%) and EV71 (45.15%), and the proportion of other enteroviruses increased significantly since 2011. CA16 strains isolated in this area have high homology, and most of the isolates in mainland China in recent years formed a phylogenetic tree closer genetic group, there was no significant change in virulence.