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Epidemiology and genetic characteristics of coxsackievirus a16 associated with hand-foot-and-mouth disease in Yantai city,China in 2018-2021

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生物安全与健康(英文) 2023年第3期5卷 181-186页

作者： Zhenlu Sun Ji Wang Xin Chi Peihua Niu Ruiqing Zhang Qiao Gao Yong Zhang Xuejun Ma Yantai Center for Disease Control and Prevention Yantai 264003China NHC Key Laboratory of Medical Virology and Viral Diseases National Institute for Viral Disease Control and PreventionChinese Center for Disease Control and PreventionBeijing 102206China WHO WPRO Regional Polio Reference Laboratory NHC Key Laboratory for BiosafetyNHC Key Laboratory for Medical VirologyNational Institute for Viral Disease Control and PreventionChinese Center for Disease Control and PreventionBeijing 102206China Center for Biosafety Mega-science Chinese Academy of ScienceWuhan 430071China

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.

关键词： Hand-foot-and-mouthdisease coxsackievirus a16 System development Genotype Phylogenetic analysis

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An open conformation determined by a structural switch for 2A protease from coxsackievirus a16

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Protein & Cell 2013年第10期4卷 782-792页

作者： Yao Sun Xiangxi Wang Shuai Yuan Minghao Dang Xuemei Li Xuejun CZhang Zihe Rao National Laboratory of Biomacromolecules Institute of BiophysicsChinese Academy of SciencesBeijing 100101China University of Chinese Academy of Sciences Beijing 100049China

coxsackievirus a16 belongs to the family Picornaviridae,and is a major agent of hand-foot-and-mouth disease that infects mostly children,and to date no vaccines or antivi-ral therapies are available.2A protease of enterovirus is a nonstructural protein and possesses both self-cleavage activity and the ability to cleave the eukaryotic translation initiation factor 4G.Here we present the crystal structure of coxsackievirus a162A protease,which interestingly forms hexamers in crystal as well as in solution.This structure shows an open conformation,with its active site accessible,ready for substrate binding and cleav-age activity.In conjunction with a previously reported“closed”state structure of human rhinovirus 2,we were able to develop a detailed hypothesis for the conforma-tional conversion triggered by two“switcher”residues Glu88 and Tyr89 located within the bll2-cII loop.Substrate recognition assays revealed that amino acid residues P1′,P2 and P4 are essential for substrate specificity,which was verifi ed by our substrate binding model.In addition,we compared the in vitro cleavage effi ciency of 2A pro-teases from coxsackievirus a16 and enterovirus 71 upon the same substrates by fl uorescence resonance energy transfer(FRET),and observed higher protease activity of enterovirus 71 compared to that of coxsackievirus a16.In conclusion,our study shows an open conformation of coxsackievirus a162A protease and the underlying mechanisms for conformational conversion and substrate specifi city.These new insights should facilitate the future rational design of effi cient 2A protease inhibitors.

关键词： coxsackievirus a16 2A protease crystal structure switcher

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Molecular epidemiology of coxsackievirus a16 circulating in children in Beijing, China from 2010 to 2019

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World Journal of Pediatrics 2021年第5期17卷 508-516页

作者： Ya-Fang Hu Li-Ping Jia Fang-Yuan Yu Li-Ying Liu Qin-Wei Song Hui-Jin Dong Jie Deng Yuan Qian Lin-Qing Zhao Li Deng Hui Huang Ru-Nan Zhu Laboratory of Virology.Beijing Key Laboratory of Etiology of Viral Diseases in Children Capital Institute of Pediatrics2 Yabao RoadChaoyang DistrictBeijing 100020China Department of Clinical Laboratory Shanghai Children's HospitalShanghai Jiaotong UniversityLuding Road 355Putuo DistrictShanghai 200040China Department of Clinical Laboratory Children's Hospital of Capital Institute of Pediatrics2 Yabao RoadBeijing 100020China Department of Infectious Diseases Children's Hospital of Capital Institute of Pediatrics2 Yabao RoadBeijing 100020China

Background coxsackievirus a16(CVa16)is one of the major etiological agents of hand,foot and mouth discase(HFMD).This study aimed to investigate the molecular epidemiology and evolutionary characteristics of CVa16.Methods Throat swabs were collected from children with HFMD and suspected HFMD during 2010-2019.Enteroviruses(EVs)were detected and typed by real-ime reverse transcription-polymerase chain reaction(RT-PCR)and RT-PCR.The genotype,evolutionary rate,the most recent common ancestor,population dynamics and selection pressure of CVa16 were analyzed based on viral protein gene(VPI)by bioinformatics software.Results A total of 4709 throat swabs were screened.EVs were detected in 3180 samples and 814 were CVa16 positive.More than 81%of CVA 16-positive children were under 5 years old.The prevalence of CVA 16 showed obvious periodic fluctuations with a high level during 2010--2012 followed by an apparent decline during 2013--2017.However,the activities of CVa16 increased gradually during 2018-2019.All the Beijing CVa16 strains belonged to sub-genotype BI,and B Ib was the dominant strain.One B Ic strain was detected in Bejing for the first time in 2016.The estimated mean evolutionary rate of VPI gene was 4.49x 103 substitution/site/year.Methionine gradually fixed at site-23 of VP1 since 2012.Two sites were detected under episodic positive selection,one of which(site-223)located in neutralizing linear epitope PEP71.Conclusions The dominant strains of CVA 16 belonged to clade B lb and evolved in a fast evolutionary rate during 2010-2019 in Beiing.To provide more favorable data for HFMD prevention and control,it is necessary to keep attention on molecular epidemiological and evolutionary characteristics of CVa16.

关键词： coxsackievirus a16 Genetic evolution Molecular epidemiology Phylogenetic analysis

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Molecular Epidemiology of Human coxsackievirus a16 Strains in Jiuiiang Region of China in 2012

Molecular Epidemiology of Human coxsackievirus A16 Strains i...

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第九届全国免疫学学术大会

作者： Wen min Yu Huanxin Xu Jin Peng Medical school of Southeast University The Third People's Hospital of Jiujiang Affiliated Hospital of Jiujiang University

Background:The hand,foot and mouth disease(HFMD)epidemics were mainly caused by human enterovirus71(EV71)and/or human coxsackievirus a16(Ca16).Studies have mainly focused on EV71 but not Ca16,because the most severe or fatal cases were caused by EV71.however,many recent reports show that Ca16 infections might lead to severe health issues.Object The present study focuses on the molecular epidemiology of human coxsackievirus a16(Ca16)strains in Jiujiang region of China in 2012.Methods:1151 specimens(throat swabs)were collected from 1151 patients with HFMD symptoms in The Third Peo-pie’s Hospital of Jiujiang City.Viral RNAs isolated from these specimens were tested by real time RT-PCR by Detection Kit.The VP1 sequence of the 6 strains of Ca16 were amplified using the Qiagen One-Step RT-PCR Kit with F:5’-CATTATTACAATGCCTACCAC-3’and R:5’-TCAGTCCCTACTGTCCTAATG-3’.The products were cloned into pMD19-T vector,subsequently sequenced and aligned.constructed the phylogenetic tree using the neighbor-joining algorithm with1000 bootstrap replicates.Results:Of all the 1151 samples,EV71 was the most prevalent agent with 50.82%(585/1151)in all samples,followed by Ca16 in 19.63%(226/1151)samples.However,Of all the positive cases,36.1%(292/811)subjects developed complications.45 patients(15.4%,44/292)infected by Ca16 developed complications such as fever and upper respiratory tract symptoms(cough,runny nose,rapid breathing).These sequences were compared using the 378bp region of VP1(2785-3162 nt,reference U05876).The genotype analysis revealed that the Ca16 strains of the Jiujiang region belonged to the B1b subgenotype and be genetically similar to the B1b strains of Ca16 prevalent in Shanghai.Conclusion:B1b subgenotype of the Ca16 was the prevalent agent of HFMD in children of the Jiujiang region.

关键词： HFMD epidemiology coxsackievirus a16

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Characterization and molecular phylogeny of coxsackievirus a16 strains associated with hand,foot,and mouth disease epidemics in Hubei province of China,in 2010 and 2011

Characterization and molecular phylogeny of coxsackievirus A...

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2012全国临床微生物与感染免疫学术研讨会

作者： Liu Wanhong~1,Dong Lanlan~1,Shi Yingying~1,Liu Yongjuan~1,Zhou Bingfei~1,Fu Chong~1,He Xiaohua~(1,2) 1.School of Basic Medical Sciences,Wuhan University 2.Wuhan Center for Medical Treatment

coxsackievirus a16(Ca16),a RNA virus of the family Picornaviridae,is cocirculating with Human enterovirus 71(HEV71) to cause hand,foot,and mouth disease(HFMD) worldwide.To clarify the characterization and molecular phylogeny of Ca16 strains associated with HFMD epidemics in Hubei province of China, throat swab specimens from 205 cases were collected during 2010-2011.78 specimens were collected from April to May,2010 and 127 specimens were collected from April to August,2011.Reverse transcriptase-polymerase chain reaction(PCR) followed by sequencing and phylogenetic analysis served to detect enterovirus and determine subtype.Enterovirus was found in 29.48%(23/78) and 29.92%(38/127) of cases and coxsackievirus a16 in 7.69%(6/78) and 11.02%(14/127) of laboratory-confirmed cases in 2010 and 2011,respectively.Phylogenetic analysis of the sequence of the VPl region of isolates indicated that most of the Hubei strains belong to the C1 subgenotype except one strain from Enshi city belongs to C3.This finding suggests that C1 is a major subgenotype of Ca16 that caused HFMD in Hubei province and additional surveilance of epidemiology and monitoring of disease severity should be continued.

关键词： coxsackievirus a16 HFMD Molecular phylogeny VP1

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Serum cholinesterase: a potential assistant biomarker for hand, foot, and mouth disease caused by enterovirus 71 infection

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Infectious Diseases of Poverty 2016年第1期5卷 249-253页

作者： Bang-Ning Cheng Yu-Lian Jin Bi-Quan Chen Li-Yan Zhu Zi-Cheng Xu Tao Shen Clinical Laboratory Center Anhui Provincial Children’s HospitalHefei 230051China Department of Infectious Diseases Anhui Provincial Children’s HospitalHefei 230051China Department of Microbiology and Infectious Disease Center School of Basic Medical SciencesPeking UniversityBeijing 100191China

Background:Hand,foot,and mouth disease(HFMD)caused by enterovirus 71(EV71)is a potentially life-threatening infectious disease that commonly occurs in children.Diagnosis of HFMD caused by EV71 largely depends on clinical manifestations and rare serological biomarkers used to identify children suffering from HFMD.Serum cholinesterase(SChE)activity has frequently been reported as a potential biomarker for solid central nervous system tumors,chronic heart failure,and liver cirrhosis.However,its potential value in the diagnosis of neurotropic virus infections,such as HFMD caused by EV71,remains to be determined.Findings:In our study,220 children hospitalized with HFMD caused by EV71,34 inpatients infected with coxsackievirus a16(CVa16),and 43 undefined enterovirus-infected HFMD inpatients were recruited at the Anhui Provincial Children’s Hospital between January 2011 and December 2012.SChE activity was measured.The non-parametric Mann–Whitney U test showed that SChE activity in children diagnosed with HFMD caused by EV71 was significantly higher than in healthy controls(p0.05).An important finding was that SChE activity declined in the recovery phase of HFMD caused by EV71 compared to the acute phase(p<0.001).Conclusions:Elevated SChE activity was observed in patients with severe HFMD caused by EV71.Therefore,SChE might be a potential assistant biomarker for the diagnosis of HFMD caused by EV71 in children.

关键词： Serum cholinesterase Hand foot and mouth disease Enterovirus 71 coxsackievirus a16 Children Biochemical marker

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Identification of Cinobufagin and Resibufogenin as Inhibitors of Enterovirus 71 Infection

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Chemical Research in Chinese Universities 2014年第6期30卷 953-958页

作者： CHEN Jiawen XU Lin SUN Shiyang ZHANG Huafei MA Tonghui SU Weiheng JIANG Chunlai National Engineering Laboratory for AIDS Vaccine Key Laboratory for Molecular Enzymology & Engineering Ministry of Education School of Life Seiences Jilin University Changchun 130012 P. R. China" College of Basic Medical Sciences Dalian Medical University Dalian 116044 t~ R. China

In this paper, cinobufagin and resibufogenin were found to inhibit enterovirus 71（EV71） infection in vitro in cell viability and plaque reduction assays. The 50% inhibitory concentrations（lCs0） of einobufagin and resibufoge- nin were （10.94±2.36） and （218±31） nmol/L, respectively, the 50% cytotoxic concentrations（CCs0） of them were （1277±223） and （1385±254） nmol/L, respectively, and the anti-EV71 selectivity index（SI50） of cinobufagin was 116.7, which are promisingly developed into drug. Using a VP1 detection assay and a constructed reporter luciferase, we found that cinobufagin and resibufogenin disrupted the synthesis of EV71 protein. However, neither of them inhibited EV71 RNA replication. Our study suggests that cinobufagin and resibufogenin are the promising candidates that should he fllrther investigated for the treatment of EV71 caused disease.

关键词： Enterovirus 71 coxsackievirus a16 Cinobufagin Resibufogenin Chansu

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