研究2014年安徽省手足口病（Hand,foot and mouth disease,HFMD）患儿中分离的柯萨奇病毒A组16型（Coxsachivirus A 16,CVA16）毒株VP1区基因特征。收集安徽省2014年1月至11月期间413份HFMD患儿咽拭子标本接种敏感细胞分离肠道病毒,用...研究2014年安徽省手足口病（Hand,foot and mouth disease,HFMD）患儿中分离的柯萨奇病毒A组16型（Coxsachivirus A 16,CVA16）毒株VP1区基因特征。收集安徽省2014年1月至11月期间413份HFMD患儿咽拭子标本接种敏感细胞分离肠道病毒,用荧光定量RT-PCR鉴定细胞培养物,对CVA16阳性培养物进行毒株VP1区RT-PCR扩增及核苷酸序列测定和基因特征分析,并与国内外参考序列构建基因亲缘性关系树。共分离鉴定出肠道病毒阳性培养物97份,其中CVA16病毒分离株17株,人肠道病毒71型（Human enterovirus 71,HEV71）分离株76株,其它肠道病毒分离株4株,总体病毒分离率为23.49%（97/413）。CVA16基因亲缘性关系分析显示：安徽省2014分离的17株CVA16毒株都属于B1基因型B1b一个分支,它们之间在核苷酸和氨基酸水平上的同源性分布为分别为95.30%~100%和98.70%~100%,但在B1b分支内形成几个传播链。17株CVA16毒株VP1区核苷酸序列与国内云南、湖南、广东、西藏和江苏地区病毒分离株同源性较高,亲缘性关系近,其中与湖南2013年和广东深圳2014年CVA16病毒分离株同源性最高,为96.40%~99.70%。2014年安徽省分离的CVA16病毒分离株属于B1基因型B1b亚型,为优势流行株;在B1b分支内形成多个小的病毒传播链共同流行。展开更多
目的比较手足口病门诊患者疱疹液、咽拭液肠道病毒检出率及一致性,分析病例群流行病学特征。方法以准分层随机法抽取同人疱疹液和咽拭液配对标本451人份,用实时荧光定量逆转录多聚酶链反应检测柯萨奇病毒A组16型(coxsackievirus A group...目的比较手足口病门诊患者疱疹液、咽拭液肠道病毒检出率及一致性,分析病例群流行病学特征。方法以准分层随机法抽取同人疱疹液和咽拭液配对标本451人份,用实时荧光定量逆转录多聚酶链反应检测柯萨奇病毒A组16型(coxsackievirus A group 16 strain,CVA16)、肠道病毒71型(enterovirus71,EV71)和总肠道病毒(Pan-enterovirus,EV)。统计分析用卡方检验、Kappa检验、秩和检验。结果疱疹液标本EV、CVA16、EV71阳性率为77.38%、39.47%、28.16%,咽拭液标本为64.97%、31.49%、26.39%,疱疹液检出率为高(χ^2值为39.20,24.92,3.06, P 值为0.000,0.000,0.077)。EV71、CVA16、EV检出一致性的Kappa检验K值为0.911,0.750,0.576, P<0.001,差异有统计学意义。疱疹液、咽拭液中EV、CVA16、EV71检出率顺序,与其Kappa检验K值顺序相反。病例星期分布性别比稳定,动态趋势一致但发病不均匀,呈以星期六为顶点并递降的峰形。结论手足口病门诊标本疱疹液优于咽拭液;病原检出率顺序与检出一致性K值顺序不一致;在一般流行病学特征下,手足口病发病星期分布可能不均并伴跨周单峰;2016年十堰市手足口病主要病原是 CVA16、EV71。展开更多
Coxsackievirus A16(CVA16),together with enterovirus type 71(EV71),is responsible for most cases of hand,foot and mouth disease(HFMD) worldwide.Recent findings suggest that the recombination between CVA16 and EV71,and ...Coxsackievirus A16(CVA16),together with enterovirus type 71(EV71),is responsible for most cases of hand,foot and mouth disease(HFMD) worldwide.Recent findings suggest that the recombination between CVA16 and EV71,and the co-circulation of these two viruses may have contributed to the increase of HFMD cases in China over the past few years.It is therefore important to further understand the virology,epidemiology,virus-host interactions and host pathogenesis of CVA16.In this study,we describe the viral kinetics of CVA16 in human rhabdomyosarcoma(RD) cells by analyzing the cytopathic effect(CPE),viral RNA replication,viral protein expression,viral RNA package and viral particle secretion in RD cells.We show that CVA16 appears to first attach,uncoat and enter into the host cell after adsorption for 1 h.Later on,CVA16 undergoes rapid replication from 3 to 6 h at MOI 1 and until 9 h at MOI 0.1.At MOI 0.1,CVA16 initiates a secondary infection as the virions were secreted before 9 h p.i.CPE was observed after 12 h p.i.,and viral antigen was first detected at 6 h p.i.at MOI 1 and at 9 h p.i.at MOI 0.1.Thus,our study provides important information for further investigation of CVA16 in order to better understand and ultimately control infections with this virus.展开更多
文摘研究2014年安徽省手足口病（Hand,foot and mouth disease,HFMD）患儿中分离的柯萨奇病毒A组16型（Coxsachivirus A 16,CVA16）毒株VP1区基因特征。收集安徽省2014年1月至11月期间413份HFMD患儿咽拭子标本接种敏感细胞分离肠道病毒,用荧光定量RT-PCR鉴定细胞培养物,对CVA16阳性培养物进行毒株VP1区RT-PCR扩增及核苷酸序列测定和基因特征分析,并与国内外参考序列构建基因亲缘性关系树。共分离鉴定出肠道病毒阳性培养物97份,其中CVA16病毒分离株17株,人肠道病毒71型（Human enterovirus 71,HEV71）分离株76株,其它肠道病毒分离株4株,总体病毒分离率为23.49%（97/413）。CVA16基因亲缘性关系分析显示：安徽省2014分离的17株CVA16毒株都属于B1基因型B1b一个分支,它们之间在核苷酸和氨基酸水平上的同源性分布为分别为95.30%~100%和98.70%~100%,但在B1b分支内形成几个传播链。17株CVA16毒株VP1区核苷酸序列与国内云南、湖南、广东、西藏和江苏地区病毒分离株同源性较高,亲缘性关系近,其中与湖南2013年和广东深圳2014年CVA16病毒分离株同源性最高,为96.40%~99.70%。2014年安徽省分离的CVA16病毒分离株属于B1基因型B1b亚型,为优势流行株;在B1b分支内形成多个小的病毒传播链共同流行。
文摘目的比较手足口病门诊患者疱疹液、咽拭液肠道病毒检出率及一致性,分析病例群流行病学特征。方法以准分层随机法抽取同人疱疹液和咽拭液配对标本451人份,用实时荧光定量逆转录多聚酶链反应检测柯萨奇病毒A组16型(coxsackievirus A group 16 strain,CVA16)、肠道病毒71型(enterovirus71,EV71)和总肠道病毒(Pan-enterovirus,EV)。统计分析用卡方检验、Kappa检验、秩和检验。结果疱疹液标本EV、CVA16、EV71阳性率为77.38%、39.47%、28.16%,咽拭液标本为64.97%、31.49%、26.39%,疱疹液检出率为高(χ^2值为39.20,24.92,3.06, P 值为0.000,0.000,0.077)。EV71、CVA16、EV检出一致性的Kappa检验K值为0.911,0.750,0.576, P<0.001,差异有统计学意义。疱疹液、咽拭液中EV、CVA16、EV71检出率顺序,与其Kappa检验K值顺序相反。病例星期分布性别比稳定,动态趋势一致但发病不均匀,呈以星期六为顶点并递降的峰形。结论手足口病门诊标本疱疹液优于咽拭液;病原检出率顺序与检出一致性K值顺序不一致;在一般流行病学特征下,手足口病发病星期分布可能不均并伴跨周单峰;2016年十堰市手足口病主要病原是 CVA16、EV71。
基金Partly supported by the National Natural Science Foundation of China (No. 20872048)
文摘Coxsackievirus A16(CVA16),together with enterovirus type 71(EV71),is responsible for most cases of hand,foot and mouth disease(HFMD) worldwide.Recent findings suggest that the recombination between CVA16 and EV71,and the co-circulation of these two viruses may have contributed to the increase of HFMD cases in China over the past few years.It is therefore important to further understand the virology,epidemiology,virus-host interactions and host pathogenesis of CVA16.In this study,we describe the viral kinetics of CVA16 in human rhabdomyosarcoma(RD) cells by analyzing the cytopathic effect(CPE),viral RNA replication,viral protein expression,viral RNA package and viral particle secretion in RD cells.We show that CVA16 appears to first attach,uncoat and enter into the host cell after adsorption for 1 h.Later on,CVA16 undergoes rapid replication from 3 to 6 h at MOI 1 and until 9 h at MOI 0.1.At MOI 0.1,CVA16 initiates a secondary infection as the virions were secreted before 9 h p.i.CPE was observed after 12 h p.i.,and viral antigen was first detected at 6 h p.i.at MOI 1 and at 9 h p.i.at MOI 0.1.Thus,our study provides important information for further investigation of CVA16 in order to better understand and ultimately control infections with this virus.
