【摘要】 目的对引起聊城地区手足口病(HFMD)流行的肠道病毒CVA16进行生物学分析,掌握其基因特征,为聊城地区手足口病的防控提供病原学资料。方法对2010~2013年聊城市各辖区采集手足口病患儿标本采用Real-time PCR法检测肠道病毒CVA16,用人横纹肌瘤(RD)细胞对核酸检测阳性标本进行病毒分离,对分离到的CVA16病毒分离株采用RT-PCR方法进行VP1区基因扩增,并对扩增产物进行序列测定,利用DNAStar和MEGA 4.0分析软件进行同源性分析和构建系统进化树。结果 2010~2013年聊城市各辖区共送检手足口病标本862份,639份标本检测为通用肠道病毒核酸阳性,其中CVA16阳性标本181份,分离到109株毒株;对毒株进行基因序列分析显示,2010~2013年聊城市流行的CVA16病毒株为B1基因型,2010和2011年为B1a和B1b两种基因亚型共循环,2012和2013年为B1b型,不同毒株VP1区基因序列在核苷酸和氨基酸水平上同源性分别为91.0%~97.0%和98.3%~100%,与近源的亚型代表株相比,聊城市优势毒株的氨基酸变异一般发生在VP1第23位(L→M)。结论聊城市2010~2013年手足口病CVA16流行毒株为B1基因型,存在B1a、B1b两个基因亚型,同源性分析表明辖区部分CVA16分离株B1a亚型与我国台湾及周边国家的分离株关系较近,B1b亚型与来自国内北方分离株形成进化树上关系较近的一群。聊城市存在不同的传播链,流行的CVA16相对保守,但已出现一定程度的变异。更多还原

【Abstract】 Objective To analyze the biological features of Coxsachie virus A16(CVA16)which caused Hand-foot-mouth Disease(HFMD)epidemics in Liaocheng area,master its gene characteristics,so as to provide the etiological data for prevention and control of HFMD. Methods Specimens from HFMD cases were collected in Liaocheng area during 2010-2013 and detected for CVA16 virus by Real-time PCR.Positive specimens were separated by using human rhabdomyosarcoma(RD)cells for virus isolation.The virus isolates were amplified and identified using RT-PCR for VP1 gene amplification.The amplified products were sent to be sequenced and the homology analysis and the construction of phylogenetic trees of CVA16 isolates were carried on by DNAStar and MEGA 4.0. Results A total of 862 specimens were collected and tested for enterovirus RNA during 2010-2013 in Liaocheng area,and 639 specimens were positive for universal primers for enterovirus.181 specimens were CVA16 positive,and of which 109 virus isolates were separated.According to the gene analysis,genotype B1 of CVA16was prevalent during 2010-2013.The gene types of B1 aand B1bwere both circulated and prevalent in 2010 and 2011,B1 bwas prevalent during 2012-2013.The homology analysis on nucleotide and amino acid for VP1 region of CVA16 virus isolates from Liaocheng area illustrated that the gene homologies were respectively 91.0%-97.0% and 98.3%-100%.Compared with the proximal subtype of representative strains,the variation on amino acid for VP1 region of CVA16 virus isolates from Liaocheng area usually occurred in the VP1 23rd(L→M). Conclusion B1 genotype is dominant among CVA16 isolates during 2010- 2013 in Liaocheng area including B1 aand B1btwo sub- types.Homology analysis indicates that partial B1 asubtypes have a close relationship with isolates from Taiwan province in China and neighboring countries.B1 bsubtypes have a close relationship with isolates from northern China and constitute a same group in the phylogenetic tree.There are different transmissions of HFMD in Liaocheng area.CVA16 isolates are relatively conservative,but there has been a certain degree of variation.更多还原