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
A sensitive reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for human enterovirus 71 (EV71) and Coxsackievirus A16 (CVA16) infection was further evaluated. The one step reaction was perfor...A sensitive reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for human enterovirus 71 (EV71) and Coxsackievirus A16 (CVA16) infection was further evaluated. The one step reaction was performed in a single tube at 65?C for 45 min for EV71 and 35 min for CVA16. The detection limits of RT-LAMP assays for both EV71 and CVA16 were 0.1 of a 50% tissue culture infective dose (TCID50) per reaction, based on 10—Fold dilutions of a titrated EV71 or CVA16 strain. The specific assay showed there were no cross-reactions with Coxsackievirus A (CVA) viruses (CVA 2, 4, 5, 7, 9, 10, 14, and 25), Coxsackievirus B (CVB) viruses (CVB 1, 2, 3, 4, and 5) or ECHO viruses (ECHO 3, 6, 11, and 19). In parallel with commercial quantitative real-time polymerase chain reaction (qRT-PCR) diagnostic kits for EV71 and CVA16, the RT-LAMP assay was evaluated with 515 clinical specimens, the results showed the RT-LAMP assay and the qRT-PCR assay were in complete agreement for 513/515 (99.6%) of the specimens. Two samples with discrepant results from two methods were further verified by nested reverse transcription polymerase chain reaction (nRT-PCR) assay and sequencing to be true positives for CVA16. In conclusion, RT-LAMP assay is demonstrated to be a sensitive and specific assay and have a great potential for the rapid and visual screening of EV71 and CVA16 in China, especially in those resource-limited hospitals and rural clinics of provincial and municipal regions.展开更多
目的:对比分析15~24个月及2~5岁CA16感染的普通型与危重症手足口病(hand,foot and mouth disease,HFMD)患者外周血中CD8^+T细胞的表达情况,寻找发现与CA16致病有关的免疫病理因素。方法:收集2014年5-8月间昆明市儿童医院感染科确诊的儿...目的:对比分析15~24个月及2~5岁CA16感染的普通型与危重症手足口病(hand,foot and mouth disease,HFMD)患者外周血中CD8^+T细胞的表达情况,寻找发现与CA16致病有关的免疫病理因素。方法:收集2014年5-8月间昆明市儿童医院感染科确诊的儿童HFMD病例外周血标本共72例,其中普通型32例,危重症40例,采用流式细胞术对患者外周血CD8^+T淋巴细胞亚群进行检测分析。结果:15~24个月普通型HFMD患者外周血CD8^+T淋巴细胞百分比略高于正常健康儿童参考值,而危重症患者CD8^+T细胞略低于正常参考值。此外,与正常参考值相比,2~5岁普通型及危重症患者外周血中CD8^+T淋巴细胞百分比均减低,其中危重症患者略低于普通型患者。结论:CA16感染后,不同年龄、不同病情的HFMD患者外周血中CD8^+T细胞的表达变化不太明显,说明CA16感染后,患者体内的CD8^+T细胞基本能够发挥正常的抗病毒免疫效应。而在危重症时,两个年龄段患者CD8^+T细胞百分比略低或减低,说明CA16的持续性感染可能对机体CD8^+T细胞的表达产生了影响,使其杀伤病毒效应减低,这可能是CA16感染诱导神经系统并发症发生的免疫病理因素之一。展开更多
In 2008,China launched a national surveillance system for hand‐foot‐and‐mouth disease(HFMD).Several million cases of HFMD are reported every year,coxsackievirus A16(CVA16)was the leading cause of HFMD epidemic in Y...In 2008,China launched a national surveillance system for hand‐foot‐and‐mouth disease(HFMD).Several million cases of HFMD are reported every year,coxsackievirus A16(CVA16)was the leading cause of HFMD epidemic in Yantai city,China in recent years,but the information of epidemiology and molecular characterization of CVA16 in Yantai is limited.The aim of this study is to investigate the epidemiological characteristics and pathogenic spectrum of HFMD,and most importantly,the molecular characterization of CVA16 in Yantai from 2018 to 2021.A total of 2,000 clinical samples were collected in Yantai city from 2018 to 2021 and the enterovirus typing was performed using real‐time reverse transcriptase–polymerase chain reaction(qRT‐PCR).VP1 coding regions of 41 CVA16 isolates were amplified and Sanger sequenced,and phylogenetic analysis was performed.During the study period,HFMD became prevalent from May to August each year.It peaked in June and declined in September.The incidence was highest in children aged 1 to 5 years,while more common in males than females.1,617 out of 2,000 clinical collection of samples were tested positive for enterovirus.Among them,614 were identified as CVA16,45 were enterovirus A71(EV A17),and 958 were other enterovirus serotypes.All 41 CVA16 strains belonged to the Bla and B1b genotypes.Homology analysis showed that 41 CVA16 isolates shared 83.2%–100%nucleotide and 93.7%–100%amino acid similarity among themselves.The results of this study update molecular epidemiology of CVA16 and provide a reference for HFMD prevention and control.展开更多
A组16型柯萨奇病毒(Coxsackievirus A16,CVA16)是引起手足口病(Hand,foot and mouth disease,HFMD)的主要病原体之一。近年来,HFMD在亚太地区暴发流行,已经成为重大的公共卫生问题之一。加强对CVA16生物学特征、流行病学特征、临床表现...A组16型柯萨奇病毒(Coxsackievirus A16,CVA16)是引起手足口病(Hand,foot and mouth disease,HFMD)的主要病原体之一。近年来,HFMD在亚太地区暴发流行,已经成为重大的公共卫生问题之一。加强对CVA16生物学特征、流行病学特征、临床表现、实验室诊断、防治手段的认识,有助于防控HFMD的蔓延。展开更多
柯萨奇病毒A组16型(Coxsackievirus A group 16 strain,CA16)感染在中国很常见,CA16在流行过程中不断发生变异,其致病性与肠道病毒71型有明显差异。CA16可诱导产生多种微小核糖核酸,影响炎性信号通路的传导,导致病理损害的发生。目前已...柯萨奇病毒A组16型(Coxsackievirus A group 16 strain,CA16)感染在中国很常见,CA16在流行过程中不断发生变异,其致病性与肠道病毒71型有明显差异。CA16可诱导产生多种微小核糖核酸,影响炎性信号通路的传导,导致病理损害的发生。目前已成功建立了小鼠、树鼩、猕猴CA16感染模型,可用于评估抗CA16药物和疫苗效果。检测方面,多重实时逆转录聚合酶链反应可针对手足口病进行高灵敏度检测和快速分型,胶体金免疫层析法可用于现场的快速诊断。多种单价、多价的CA16疫苗在动物实验中诱导出有效的免疫。展开更多
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.
文摘A sensitive reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for human enterovirus 71 (EV71) and Coxsackievirus A16 (CVA16) infection was further evaluated. The one step reaction was performed in a single tube at 65?C for 45 min for EV71 and 35 min for CVA16. The detection limits of RT-LAMP assays for both EV71 and CVA16 were 0.1 of a 50% tissue culture infective dose (TCID50) per reaction, based on 10—Fold dilutions of a titrated EV71 or CVA16 strain. The specific assay showed there were no cross-reactions with Coxsackievirus A (CVA) viruses (CVA 2, 4, 5, 7, 9, 10, 14, and 25), Coxsackievirus B (CVB) viruses (CVB 1, 2, 3, 4, and 5) or ECHO viruses (ECHO 3, 6, 11, and 19). In parallel with commercial quantitative real-time polymerase chain reaction (qRT-PCR) diagnostic kits for EV71 and CVA16, the RT-LAMP assay was evaluated with 515 clinical specimens, the results showed the RT-LAMP assay and the qRT-PCR assay were in complete agreement for 513/515 (99.6%) of the specimens. Two samples with discrepant results from two methods were further verified by nested reverse transcription polymerase chain reaction (nRT-PCR) assay and sequencing to be true positives for CVA16. In conclusion, RT-LAMP assay is demonstrated to be a sensitive and specific assay and have a great potential for the rapid and visual screening of EV71 and CVA16 in China, especially in those resource-limited hospitals and rural clinics of provincial and municipal regions.
文摘目的:对比分析15~24个月及2~5岁CA16感染的普通型与危重症手足口病(hand,foot and mouth disease,HFMD)患者外周血中CD8^+T细胞的表达情况,寻找发现与CA16致病有关的免疫病理因素。方法:收集2014年5-8月间昆明市儿童医院感染科确诊的儿童HFMD病例外周血标本共72例,其中普通型32例,危重症40例,采用流式细胞术对患者外周血CD8^+T淋巴细胞亚群进行检测分析。结果:15~24个月普通型HFMD患者外周血CD8^+T淋巴细胞百分比略高于正常健康儿童参考值,而危重症患者CD8^+T细胞略低于正常参考值。此外,与正常参考值相比,2~5岁普通型及危重症患者外周血中CD8^+T淋巴细胞百分比均减低,其中危重症患者略低于普通型患者。结论:CA16感染后,不同年龄、不同病情的HFMD患者外周血中CD8^+T细胞的表达变化不太明显,说明CA16感染后,患者体内的CD8^+T细胞基本能够发挥正常的抗病毒免疫效应。而在危重症时,两个年龄段患者CD8^+T细胞百分比略低或减低,说明CA16的持续性感染可能对机体CD8^+T细胞的表达产生了影响,使其杀伤病毒效应减低,这可能是CA16感染诱导神经系统并发症发生的免疫病理因素之一。
基金supported by Shandong Provincial Preventive Medicine Association Project(LYH 2017‐26).
文摘In 2008,China launched a national surveillance system for hand‐foot‐and‐mouth disease(HFMD).Several million cases of HFMD are reported every year,coxsackievirus A16(CVA16)was the leading cause of HFMD epidemic in Yantai city,China in recent years,but the information of epidemiology and molecular characterization of CVA16 in Yantai is limited.The aim of this study is to investigate the epidemiological characteristics and pathogenic spectrum of HFMD,and most importantly,the molecular characterization of CVA16 in Yantai from 2018 to 2021.A total of 2,000 clinical samples were collected in Yantai city from 2018 to 2021 and the enterovirus typing was performed using real‐time reverse transcriptase–polymerase chain reaction(qRT‐PCR).VP1 coding regions of 41 CVA16 isolates were amplified and Sanger sequenced,and phylogenetic analysis was performed.During the study period,HFMD became prevalent from May to August each year.It peaked in June and declined in September.The incidence was highest in children aged 1 to 5 years,while more common in males than females.1,617 out of 2,000 clinical collection of samples were tested positive for enterovirus.Among them,614 were identified as CVA16,45 were enterovirus A71(EV A17),and 958 were other enterovirus serotypes.All 41 CVA16 strains belonged to the Bla and B1b genotypes.Homology analysis showed that 41 CVA16 isolates shared 83.2%–100%nucleotide and 93.7%–100%amino acid similarity among themselves.The results of this study update molecular epidemiology of CVA16 and provide a reference for HFMD prevention and control.
文摘A组16型柯萨奇病毒(Coxsackievirus A16,CVA16)是引起手足口病(Hand,foot and mouth disease,HFMD)的主要病原体之一。近年来,HFMD在亚太地区暴发流行,已经成为重大的公共卫生问题之一。加强对CVA16生物学特征、流行病学特征、临床表现、实验室诊断、防治手段的认识,有助于防控HFMD的蔓延。
文摘柯萨奇病毒A组16型(Coxsackievirus A group 16 strain,CA16)感染在中国很常见,CA16在流行过程中不断发生变异,其致病性与肠道病毒71型有明显差异。CA16可诱导产生多种微小核糖核酸,影响炎性信号通路的传导,导致病理损害的发生。目前已成功建立了小鼠、树鼩、猕猴CA16感染模型,可用于评估抗CA16药物和疫苗效果。检测方面,多重实时逆转录聚合酶链反应可针对手足口病进行高灵敏度检测和快速分型,胶体金免疫层析法可用于现场的快速诊断。多种单价、多价的CA16疫苗在动物实验中诱导出有效的免疫。
