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a novel enterovirus and parechovirus multiplex one-step real-time pcr-validation and clinical experience.as the number of new enteroviruses and human parechoviruses seems ever growing, the necessity for updated diagnostics is relevant. we have updated an enterovirus assay and combined it with a previously published assay for human parechovirus resulting in a multiplex one-step rt-pcr assay. the multiplex assay was validated by analysing the sensitivity and specificity of the assay compared to the respective monoplex assays, and a good concordance was found. furthermore, the enterovirus assay was ab ...201323845901
[2009 evolution for vp4 of enterovirus 71 strains in shenzhen].to analyze the genetic evolution for the common causative agent of hand, foot and mouth disease(hfmd) that vp4 of human enterovirus 71 in shenzhen district.201323855119
seroepidemiology of human enterovirus71 and coxsackievirusa16 among children in guangdong province, china.hand, foot and mouth disease (hfmd) is a common pediatric illness. mainly induced by the enterovirus 71 and coxsackievirus a 16 infections, the frequently occurred hfmd outbreaks have become a serious public health problem in southeast asia. currently,only a few studies have investigated the human immunity to hfmd in china. in this study, we conducted a cohort study in guangdong province, china.201323855481
a study of the virulence in mice of high copying fidelity variants of human enterovirus 71.polioviruses with a g64s mutation in the 3d polymerase have enhanced replication fidelity and are attenuated in animal models. here we describe the mouse virulence properties of high replication fidelity 3d polymerase variants of human enterovirus 71 (hev71), with mutations at positions 3d-s264l, 3d-g64r or at 3d-s264l plus 3d-g64r. mouse-adapted strains (mp-g64r, mp-s264l and mp-s264l-g64r) were constructed in order to compare the virulence of the 3d polymerase variants with that of mouse-adapt ...201323856384
global transcriptomic analysis of human neuroblastoma cells in response to enterovirus type 71 infection.human enterovirus type 71 (ev71) is the major pathogen of hand-foot-and-mouth disease (hfmd) and has been associated with severe neurological disease and even death in infants and young children. the pathogenesis of ev71 infection in the human central nervous system remains unclear. in this study, human whole genome microarray was employed to perform transcriptome profiling in sh-sy5y human neuroblastoma cells infected with ev71. the results indicated that ev71 infection lead to altered expressi ...201323861741
vp2 dominated cd4+ t cell responses against enterovirus 71 and cross-reactivity against coxsackievirus a16 and polioviruses in a healthy population.enterovirus 71 (ev71)-associated hand-foot-mouth disease has become a major threat to public health in the asia-pacific region. although t cell immunity is closely correlated with clinical outcomes of ev71 infection, little is known about t cell immunity baseline against ev71 and t cell immunogenecity of ev71 ags in the population, which has restricted our understanding of immunoprotection mechanisms. in this study, we investigated the cellular immune responses against the four structural ags of ...201323863902
complete genome sequence of a human enterovirus 71 strain isolated in brunei in 2006.the complete genome sequence of a human enterovirus 71 strain isolated in brunei in 2006 was determined. phylogenetic analysis based on the complete genome sequence classified this strain into subgenogroup b5.201323868136
characterisation of enterovirus 71 replication kinetics in human colorectal cell line, ht29.hand, foot and mouth disease (hfmd), a contagious viral disease that commonly affects infants and children with blisters and flu like symptoms, is caused by a group of enteroviruses such as enterovirus 71 (ev71) and coxsackievirus a16 (ca16). however some hfmd caused by ev71 may further develop into severe neurological complications such as encephalitis and meningitis. the route of transmission was postulated that the virus transmit from one person to another through direct contact of vesicular ...201323875129
accuracy of diagnostic methods and surveillance sensitivity for human enterovirus, south korea, 1999-2011.the epidemiology of enteroviral infection in south korea during 1999-2011 chronicles nationwide outbreaks and changing detection and subtyping methods used over the 13-year period. of 14,657 patients whose samples were tested, 4,762 (32.5%) samples were positive for human enterovirus (human ev); as diagnostic methods improved, the rate of positive results increased. a seasonal trend of outbreaks was documented. genotypes enterovirus 71, echovirus 30, coxsackievirus b5, enterovirus 6, and coxsack ...201323876671
construction and characterization of an infectious cdna clone of enterovirus type 71 subgenotype c4.enterovirus 71 (ev71) is the major causative agent of hand, foot, and mouth disease and induces fatal neurological complications. in recent years, this virus has become a major threat to public health in the asia-pacific region, while no effective antiviral therapies and vaccines are currently available. in this study, we constructed and characterized for the first time an infectious full-length ev71 cdna clone derived from the shzh98 strain, which was the first subgenotype c4 strain isolated in ...201323877719
fatal case of enterovirus 71 infection and rituximab therapy, france, 2012. 201323880543
autoprocessing: an essential step for expression and purification of enterovirus 71 3c(pro) in escherichia coli.a gene encoding the 3bc of human enterovirus 71 (ev71) was cloned and inserted into a derivative of plasmid pet-32a(+) driven by t7 promoter. the expressed 3c protease (3c(pro)) autocatalytically cleaved itself from the recombinant protein trx-3bc and the mature 3c(pro) partitioned in the soluble fraction of bacterial lysate. the 13-amino-acid peptide substrates with the junction of 3b/3c were used to verify the proteolysis activity of the purified 3c(pro). the ev71 3c(pro) had a km value of 63 ...201323881322
elevated antigen-specific th2 type response is associated with the poor prognosis of hand, foot and mouth disease.the immunopathogenesis of severe hand, foot and mouth disease (hfmd) remains elusive. this study revealed that enterovirus 71 (ev71) epitope-specific cd4+ t cell responses of hfmd patients were skewed toward a th2 cytokine profile. patients that demonstrated higher levels of il-4 expression in their cd4 t cells following antigen stimulation in vitro tended to have a more prolonged period of high fevers and a longer duration of illness. thus, an increase of ev71 epitope-specific th2 type response ...201323886670
[research on human enterovirus 71 infectivity assay based on a real-time cell analysis].this research aims to evaluate the application of real - time cell assay (rtca) based on microelectronics sensor technology in the detection of hev71 induced cell lesion. growth indexes of rd cells at different stages were observed dynamically, appropriate cell concentration was selected to test hev71 infectivity and to determine the hev71 neutralizing antibody titer in serum. the traditional microplate test was used as methodology comparison and results validation at the same time. cell impedan ...201323895003
[the 2a protease of enterovirus 71 cleaves nup62 to inhibit nuclear transport].to study the impact of the enterovirus 71(ev71) on the nuclear transport mechanism,the pgfp-nls vector with nuclear location signal(nls) was constructed, rd cells transfected by the pgfp-nls vector were inoculated with the ev71 or cotransfected by ev71-2a vector. the results showed that gfp protein with nls was expressed in the cytoplasm due to the inhibition of nuclear transport. in order to further study the mechanism of the ev71 to prevent nuclear transport,nup62 was detected by western blott ...201323895008
[recombination analysis of enterovirus 71 strain isolated in guangzhou, 2009].to describe the recombination features of human enterovirus 71 strain guangzhou09 isolated in guangzhou in 2009, the complete nucleotide sequences of guangzhou09 were analyzed by various of bioinformatics software. phylogenetic analysis based on p1, p2 and p3 regions indicated that recombination occurred between ev71 and cva4. phylogenetic, similarity plot and bootscan analysis further revealed the recombination between ev71 genotype c strain shanghai-fj713317 and cva4 strain hq728260 at region ...201323905474
[the pathogenic spectrum of hand, foot and mouth disease and molecular characterizations of human enterovirus 71 in inner mongolia autonomous region in 2010].to study the pathogenic spectrum of hand, foot and mouth disease (hfmd) and the molecular characterizations of human enteroviruses 71 (hev71) isolated from the clinical specimens of hfmd patients in inner mongolia in 2010. a total of 921 clinical specimens including stools and throat swabs were collected from hfmd patients in outpatient service in inner mongolia and then viral isolation was performed, the positive viral isolates were identified by using the real-time pcr method (detecting ev, he ...201323905475
changing aetiology of hand, foot and mouth disease in linyi, china, 2009-2011.millions of hand, foot and mouth disease cases occur annually and the disease is considered a significant public health threat in china. we have focused on linyi for 3 years to analyse the aetiological spectrum and epidemic changes. the reported number of cases of hfmd in linyi declined in 2011,with the percentage of hev71 dropping to 11%. although cva16 rose to 51% in 2011, the number of cva16 cases did not change significantly compared with the previous 2 years. other types of enteroviruses we ...201423910234
development of duplex real-time rt-pcr based on taqman technology for detecting simultaneously the genome of pan-enterovirus and enterovirus 71.human enterovirus (ev) 71 is the main etiological agent of hand, foot, and mouth disease (hfmd). it is associated with neurological complications, and caused fatalities during recent outbreaks in the asia-pacific region. infections caused by ev71 could lead to many complications, ranging from brainstem encephalitis to pulmonary oedema, resulting in high mortality. in this study, a duplex real-time rt-pcr assay was developed in order to simultaneously detect pan-ev and ev71. ev71-specific primers ...201323918544
establishment of cell lines with increased susceptibility to ev71/ca16 by stable overexpression of scarb2.human enterovirus type 71 (ev71) and coxsackievirus a group type 16 (ca16) belong to human enterovirus species a of the family picornaviridae. these viruses are recognized as the major pathogens responsible for epidemics of hand-foot-mouth disease (hfmd), which presents with fever and vesicular eruptions of palms, soles of the feet or mouth. human scavenger receptor class b, member 2 (scarb2) has been identified as the receptor for both ev71 and ca16, as overexpression of scarb2 in cells can enh ...201323919614
novel cycloalkylthiophene-imine derivatives bearing benzothiazole scaffold: synthesis, characterization and antiviral activity evaluation.a series of novel cycloalkylthiophene-imine derivatives containing benzothiazole unit were designed, synthesized and evaluated for their anti-viral activities. the bio-evaluation results indicated that some of the target compounds (such as 5g, 5i, 5u) exhibited good to moderate antiviral effect on cvb5, adv7 and ev71 viruses, however, these compounds did not have inhibition activity against h1n1 virus. especially, the compounds 4c and 4d also exhibited high antiviral activities, which provide a ...201323920438
immunogenicity, safety, and immune persistence of a novel inactivated human enterovirus 71 vaccine: a phase ii, randomized, double-blind, placebo-controlled trial.vaccination is considered a top priority for the control of human enterovirus 71 (ev71) infection outbreaks.201423922377
genetic and antigenic characterization of enterovirus 71 in ho chi minh city, vietnam, 2011.enterovirus 71 (ev71) frequently causes fatal infections in young children in asia. in 2011, ev71 epidemics occurred in southern vietnam. we conducted genetic and antigenic analysis of the ev71 isolates and found that 94% of them were genotype c4a related to two lineages circulating in china and 6% were genotype c5 which have circulated in vietnam since 2003. antigenic variants were not detected. ev71 vaccines are being developed. longitudinal enterovirus surveillance data are critical to formul ...201323922846
[retrospective analysis of clinical and epidemiological characteristics of hand-foot-and-mouth disease deceased cases in zhejiang province].to understand the clinical and epidemiological characteristics of hand-foot-and-mouth disease (hfmd) deceased cases.201323927799
plasma b-type natriuretic peptide study in children with severe enterovirus 71 infection: a pilot study.severe enterovirus 71 (ev71) infections in children can result in acute heart failure. b-type natriuretic peptide (bnp) is a good biomarker of myocardial stress. the purpose of this study was to use plasma bnp for the detection of ev71 infection with cardiac involvement.201323932576
patterns of polymorphism and divergence in the vp1 gene of enterovirus 71 circulating in the asia-pacific region between 1994 and 2013.enterovirus 71 has been implicated in several outbreaks of hand, foot and mouth disease in the asia-pacific region. the present study aimed to achieve comprehensive evolutionary dynamic aspects of ev71 during 1994-2013, based on phylogenetic analyses of the vp1 sequences. the results indicated that 4 genotypes, namely c4, c1, c2 and b4 are the predominant strains, especially in southeast asian countries. no common ancestor was shared in different countries. fourteen sites of substitutions were d ...201323933074
enterovirus 71 binding to psgl-1 on leukocytes: vp1-145 acts as a molecular switch to control receptor interaction.some strains of enterovirus 71 (ev71), but not others, infect leukocytes by binding to a specific receptor molecule: the p-selectin glycoprotein ligand-1 (psgl-1). we find that a single amino acid residue within the capsid protein vp1 determines whether ev71 binds to psgl-1. examination of capsid sequences of representative ev71 strains revealed that the psgl-1-binding viruses had either a g or a q at residue 145 within the capsid protein vp1 (vp1-145g or q), whereas psgl-1-nonbinding viruses ha ...201323935488
protective efficacy of vp1-specific neutralizing antibody associated with a reduction of viral load and pro-inflammatory cytokines in human scarb2-transgenic mice.hand-foot-mouth diseases (hfmd) caused by enterovirus 71 (ev71) and coxsackievirus 16 (cva16) in children have now become a severe public health issue in the asian-pacific region. recently we have successfully developed transgenic mice expressing human scavenger receptor class b member 2 (hscarb2, a receptor of ev71 and cva16) as an animal model for evaluating the pathogenesis of enterovirus infections. in this study, hscarb2-transgenic mice were used to investigate the efficacy conferred by a p ...201323936115
[epidemiological and etiological characteristics of hand-foot-mouth disease in ningbo, zhejiang province, 2008-2011].to analyze the epidemiological characteristics of hand foot and mouth disease (hfmd) in ningbo.201323937841
soluble interleukin-6 receptor-mediated innate immune response to dna and rna viruses.the interleukin-6 (il-6) receptor, which exists as membrane-bound and soluble forms, plays critical roles in the immune response. the soluble il-6 receptor (sil6r) has been identified as a potential therapeutic target for preventing coronary heart disease. however, little is known about the role of this receptor during viral infection. in this study, we show that sil6r, but not il-6, is induced by viral infection via the cyclooxygenase-2 pathway. interestingly, sil6r, but not il-6, exhibited ext ...201323946454
a strain-specific epitope of enterovirus 71 identified by cryo-electron microscopy of the complex with fab from neutralizing antibody.enterovirus 71 (ev71) is a picornavirus that causes outbreaks of hand, foot, and mouth disease (hfmd), primarily in the asia-pacific area. unlike coxsackievirus a16, which also causes hfmd, ev71 induces severe neuropathology leading to high fatalities, especially among children under the age of 6 years. currently, no established vaccines or treatments are available against ev71 infection. the monoclonal antibody ma28-7 neutralizes only specific strains of ev71 that have a conserved glycine at am ...201323946455
transgenic mouse model for the study of enterovirus 71 neuropathogenesis.enterovirus 71 (ev71) typically causes mild hand-foot-and-mouth disease in children, but it can also cause severe neurological disease. recently, epidemic outbreaks of ev71 with significant mortality have been reported in the asia-pacific region, and ev71 infection has become a serious public health concern worldwide. however, there is little information available concerning ev71 neuropathogenesis, and no vaccines or anti-ev71 drugs have been developed. previous studies of this disease have used ...201323959904
proteomic analysis of extremely severe hand, foot and mouth disease infected by enterovirus 71.to clarify the molecular mechanisms that participate in the severe hand, foot and mouth disease (hfmd) infected by enterovirus 71 and to detect any related protein biomarkers, we performed proteomic analysis of protein extracts from 5 extremely severe hfmd children and 5 healthy children.201323961958
crystal structure of 2a proteinase from hand, foot and mouth disease virus.ev71 is responsible for several epidemics worldwide; however, the effective antiviral drug is unavailable to date. the 2a proteinase (2a(pro)) of ev71 presents a promising drug target due to its multiple roles in virus replication, inhibition of host protein synthesis and evasion of innate immunity. we determined the crystal structure of ev71 2a(pro) at 1.85å resolution, revealing that the proteinase maintains a chymotrypsin-like fold. the active site is composed of the catalytic triads c110a, h ...201323973886
coxsackievirus a6 and hand, foot and mouth disease: three case reports of familial child-to-immunocompetent adult transmission and a literature review.hand, foot and mouth disease (hfmd) is a highly contagious viral infection characterized by typical maculopapular or vesicular eruptions on the hands and feet and in the oral cavity. it affects predominantly children and/or immunocompromised adults. it usually follows a benign and self-limiting course. however, hfmd cases with severe or lethal complications such as encephalitis, meningitis, pulmonary edema and myocarditis have also been reported, mostly in children, but also in adults. high infe ...201324019771
an open conformation determined by a structural switch for 2a protease from coxsackievirus a16.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 antiviral 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 a16 2a protease, which interestingly forms hexamers in crystal as well a ...201324026848
human astrocytic cells support persistent coxsackievirus b3 infection.enteroviruses can frequently target the human central nervous system to induce a variety of neurological diseases. although enteroviruses are highly cytolytic, emerging evidence has shown that these viruses can establish persistent infections both in vivo and in vitro. here, we investigated the susceptibility of three human brain cell lines, ccf-sttg1, t98g, and sk-n-sh, to infection with three enterovirus serotypes: coxsackievirus b3 (cvb3), enterovirus 71, and coxsackievirus a9. persistent inf ...201324027313
[cloning, expression and activity determination of capsid protein vp1 of enterovirus type 71]. 201324027809
molecular epidemiology of enterovirus 71, coxsackievirus a16 and a6 associated with hand, foot and mouth disease in spain.hand, foot and mouth disease (hfmd) is a childhood illness frequently caused by genotypes belonging to the enterovirus a species, including coxsackievirus (cv)-a16 and enterovirus (ev)-71. between 2010 and 2012, several outbreaks and sporadic cases of hfmd occurred in different regions of spain. the objective of the present study was to describe the enterovirus epidemiology associated with hfmd in the country. a total of 80 patients with hfmd or atypical rash were included. detection and typing ...201424033818
[relationship between g6pd deficiency and hand-foot-mouth disease induced by enterovirus 71].to study the influence of glucose-6-phosphate dehydrogenase (g6pd) deficiency on hand-foot-mouth disease (hfmd) induced by enterovirus 71 (ev71) , and possible mechanisms.201324034918
atypical hand-foot-and-mouth disease associated with coxsackievirus a6 infection.hand-foot-and-mouth disease (hfmd) is an acute viral illness commonly caused by coxsackievirus (cv)-a16 and enterovirus 71 infections. recently, atypical hfmd has been reported in association with cv-a6, an uncommon enterovirus strain.201324035209
environmental alkylphenols modulate cytokine expression in plasmacytoid dendritic cells.alkylphenols, such as nonylphenol (np) and 4-octylphenol (4-op), have the potential to disturb immune system due to their weak estrogen-like activity, an effect with potential serious public health impact due to the worldwide distribution of these substances. plasmacytoid dendritic cells (pdcs) can secrete large amounts of type i ifns and are critical in immune regulation. however, there has been limited study about the influence of alkylphenols on the function of pdcs.201324039973
enterovirus 71 vp1 activates calmodulin-dependent protein kinase ii and results in the rearrangement of vimentin in human astrocyte cells.enterovirus 71 (ev71) is one of the main causative agents of foot, hand and mouth disease. its infection usually causes severe central nervous system diseases and complications in infected infants and young children. in the present study, we demonstrated that ev71 infection caused the rearrangement of vimentin in human astrocytoma cells. the rearranged vimentin, together with various ev71 components, formed aggresomes-like structures in the perinuclear region. electron microscopy and viral rna l ...201324073199
eupafolin and ethyl acetate fraction of kalanchoe gracilis stem extract show potent antiviral activities against enterovirus 71 and coxsackievirus a16.enterovirus 71 (ev71) and coxsackievirus a16 (coxa16) are main pathogens of hand-foot-and-mouth disease, occasionally causing aseptic meningitis and encephalitis in tropical and subtropical regions. kalanchoe gracilis, da-huan-hun, is a chinese folk medicine for treating pain and inflammation, exhibiting antioxidant and anti-inflammatory activities. our prior report (2012) cited k. gracilis leaf extract as moderately active against ev71 and coxa16. this study further rates antienteroviral potent ...201324078828
is a monovalent vaccine against enterovirus 71 sufficient? a review of enterovirus 71 vaccine development based on enterovirus surveillance in taiwan. 201324079713
bedside immunochromatographic test for enterovirus 71 infection in children.enterovirus 71 (ev71) causes frequent outbreaks worldwide, particularly in the asia-pacific area. its quick spread is a critical challenge for public health and timely preventive measures and clinical management therefore rely on early detection. there is a need for a rapid, easy-to-use, and reliable method for detecting ev71 infections.201324084600
association of functional polymorphisms in the mxa gene with susceptibility to enterovirus 71 infection.myxovirus resistance a (mxa) is an antiviral protein induced by type i interferons α and β (ifn-α and ifn-β) that can inhibit virus replication. we examined whether the mxa polymorphisms were related to the risk and severity of enterovirus 71 (ev71) infection in chinese populations. the mxa c-123a and g-88t polymorphisms were genotyped in two independent case-control populations in china by polymerase chain reaction-based restriction fragment length polymorphism (pcr-rflp) analysis. multivariate ...201424085612
heat shock protein 90: role in enterovirus 71 entry and assembly and potential target for therapy.although several factors participating in enterovirus 71 (ev71) entry and replication had been reported, the precise mechanisms associated with these events are far from clear. in the present study, we showed that heat shock protein 90 (hsp90) is a key element associated with ev71 entry and replication in a human rhabdomyosarcoma of rd cells. inhibition of hsp90 by pretreating host cells with hsp90β sirna or blocking hsp90 with a hsp90-specific antibody or geldanamycin (ga), a specific inhibitor ...201324098578
chlorogenic acid inhibits the replication and viability of enterovirus 71 in vitro.enterovirus 71 (ev71) is an etiology for a number of diseases in humans. traditional chinese herbs have been reported to be effective for treating ev71 infection. however, there is no report about the antiviral effects of cha against ev71. in this study, plaque reduction assay demonstrated that the inhibitory concentration 50% (ic50) of cha on ev71 replication is 6.3 µg/ml. when both cha (20 µg/ml) and ev71 were added, or added post-infection at different time points, cha was able to effectively ...201324098754
immunogenicity, safety, and lot consistency of a novel inactivated enterovirus 71 vaccine in chinese children aged 6 to 59 months.the determination of lot-to-lot consistency in the manufacturing process is a mandatory step in the clinical development of the novel enterovirus 71 (ev71) vaccine. a phase iii, randomized, placebo-controlled, double-blind trial assessed the lot consistency, immunogenicity, and safety of the ev71 vaccine in children aged 6 to 59 months. healthy children (n = 1,400) received one of three lots of the ev71 vaccine containing 400 u of ev71 antigen or a placebo at days 0 and 28. blood samples were co ...201324108780
human enterovirus 71 epidemics: what's next?human enterovirus 71 (ev71) epidemics have affected various countries in the past 40 years. ev71 commonly causes hand, foot and mouth disease (hfmd) in children, but can result in neurological and cardiorespiratory complications in severe cases. genotypic changes of ev71 have been observed in different places over time, with the emergence of novel genotypes or subgenotypes giving rise to serious outbreaks. since the late 1990s, intra- and inter-typic recombination events in ev71 have been increa ...201324119538
the enteroviruses: problems in need of treatments.specific antiviral therapy is currently not available for enterovirus (ev) infections. poliomyelitis, ev 71 neurologic disease, and neonatal ev disease are three manifestations of ev infections that exemplify the importance of developing antivirals for ev infections. despite tremendous strides in the effort to eradicate polio through vaccination, challenges remain, including the potential for transmission of neurovirulent vaccine-derived polioviruses which have genetically reverted from live-att ...201424119825
epidemiology of enterovirus 71 infections in taiwan.enterovirus 71 (ev71) was first described in usa in 1969 but retrospective studies in the netherlands further detected ev71 in the clinical specimens collected in 1963. ev71 has one single serotype measured by using hyperimmune animal antisera but can be phylogenetically classified into three genogroups (a, b, and c) including 11 genotypes (a, b1-b5, c1-c5). in taiwan, ev71 caused a large-scale nationwide epidemic in 1998. retrospective studies further detected ev71 in clinical specimens collect ...201424120535
chimeric virus-like particle vaccines displaying conserved enterovirus 71 epitopes elicit protective neutralizing antibodies in mice through divergent mechanisms.enterovirus 71 (ev71) is a major causative agent of hand, food, and mouth disease, which frequently occurs in young children. since there are 11 subgenotypes (a, b1 to b5, and c1 to c5) within ev71, an ev71 vaccine capable of protecting against all of these subgenotypes is desirable. we report here the vaccine potential and protective mechanism of two chimeric virus-like particles (vlps) presenting conserved neutralizing epitopes of ev71. we show that fusions of hepatitis b core antigen (hbc) wi ...201424131712
frequency of human enterovirus 71 in children under 8 years old with aseptic menengitis in tehran.human enterovirus 71 (hev71) was isolated for the first time from an infant with encephalitis in california in 1969 and then spread through the world. it has emerged as a major cause of a vast variety of diseases such as epidemics of hand, foot and mouth disease (hfmd), aseptic meningitis (am), acute flaccid paralysis, and encephalitis. the aim of this study was to determine the prevalence of enterovirus 71 in children < 8 years old who were hospitalized due to primary diagnosis of am in tehran.201324133924
evaluation of the stability of enterovirus 71 virus-like particle.enterovirus 71 (ev71) is responsible for the outbreaks of hand-foot-and-mouth disease that caused significant mortality in children, but no vaccine is available yet. ev71 virus-like particle (vlp) is the empty capsid consisting of viral structural proteins but can elicit potent immune responses, rendering vlp a promising ev71 vaccine candidate. to evaluate whether vlp remains stable after long-term storage, which is crucial for advancing the vlp vaccine to the clinical setting, we evaluated the ...201424140131
impact of endothelial nitric oxide synthase gene polymorphism on severity of enterovirus 71-infection in chinese children.genetic polymorphism g894t on the endothelial nitric oxide synthase (enos) gene has been reported as a susceptibility factor in a number of diseases, but evidence of its effect on enterovirus 71 (ev71) infection is lacking. this study investigated the possible association between this polymorphism (rs1799983) and disease severity in chinese children with ev71 infection.201324140479
enterovirus genotypes causing hand foot and mouth disease in shanghai, china: a molecular epidemiological analysis.a rapid expansion of hand, foot, and mouth disease (hfmd) outbreaks has occurred and caused deaths in china in recent years, but little is known about the other etiologic agents except enterovirus 71 (ev71) and coxsackievirus a 16 (ca16). the objective of this study is to determine the genotype compositions of enterovirus causing hfmd in shanghai and identify any associations between enterovirus types and clinical manifestations.201324148902
the population-based health effect of hand, foot and mouth disease in children in shanghai.an effective enterovirus 71 (ev71) vaccine is needed to control the annual outbreaks of hand, foot and mouth disease (hfmd) in china. adequate epidemiologic data relating to hfmd are needed to make decisions about appropriate public health interventions and implementation of the new ev71 vaccine.201424153009
enterovirus 71 pathogenicity in monkeys and cotton rats.enterovirus 71 (ev71) is a neurovirulent non-polio enterovirus that can cause severe central nervous system (cns) infection in infants. vervet monkeys infected intracerebrally or intramuscularly with ev71 isolates from the bulgarian outbreak of 1975 developed clinical manifestations and pathological signs of encephalomyelitis and spinal poliomyelitis that were similar to ev71 neuroinfection in children. in addition, vervet monkeys with encephalomyelitis had severe alterations in the choroid plex ...201424158347
a one-step dipstick assay for the on-site detection of nucleic acid.we have developed a one-step nucleic acid dipstick assay (nada) for visually detecting polymerase chain reaction (pcr) products within 3min. "one-step" means that there were no additional procedures between amplification and detection.201324161476
structure of limp-2 provides functional insights with implications for sr-bi and cd36.members of the cd36 superfamily of scavenger receptor proteins are important regulators of lipid metabolism and innate immunity. they recognize normal and modified lipoproteins, as well as pathogen-associated molecular patterns. the family consists of three members: sr-bi (which delivers cholesterol to the liver and steroidogenic organs and is a co-receptor for hepatitis c virus), limp-2/lgp85 (which mediates lysosomal delivery of β-glucocerebrosidase and serves as a receptor for enterovirus 71 ...201324162852
clinico-radiological spectrum in enterovirus 71 infection involving the central nervous system in children.enterovirus 71 infection causes hand, foot and mouth disease in children, and can produce diverse neurologic complications. epidemics occurring in korea between 2009 and 2012 resulted in the death of some patients. the present study aimed to clarify the correlation between clinical features and mri findings in patients presenting with acute neurologic manifestations related to enterovirus 71 infection. based on their clinical features, the patients were classified into four clinical groups: (1) ...201424169271
coxsackievirus a6-induced hand-foot-mouth disease.hand-foot-mouth disease (hfmd) is an acute, self-limited, highly contagious viral illness that commonly affects children younger than 5 years. it is most typically caused by enterovirus 71 or coxsackievirus a16 and results in asymptomatic infection or mild disease. immunocompetent adults are rarely affected. recently, there have been increasing reports of a more severe form of hfmd associated with fevers, joint pains, and widespread painful eruptions. some of these patients required hospitalizat ...201324172861
epidemiology and clinical characteristics of hand foot, and mouth disease in a shenzhen sentinel hospital from 2009 to 2011.we investigated the epidemiological and clinical data of all hand, foot, and mouth disease (hfmd) cases in a sentinel hospital of shenzhen, china from 2009 to 2011.201324225231
coxsackievirus a16: epidemiology, diagnosis, and vaccine.coxsackievirus 16 (ca16) is one of the major pathogens associated with hand, foot, and mouth disease (hfmd) in infants and young children. in recent years, ca16 and human enterovirus 71 (ev71) have often circulated alternatively or together in the western pacific region, which has become an important public health problem in this region. hfmd caused by ca16 infection is generally thought to be mild and self-limiting. however, recently several severe and fatal cases involving ca16 have been repor ...201424231751
modification of picornavirus genomic rna using 'click' chemistry shows that unlinking of the vpg peptide is dispensable for translation and replication of the incoming viral rna.picornaviruses constitute a large group of viruses comprising medically and economically important pathogens such as poliovirus, coxsackievirus, rhinovirus, enterovirus 71 and foot-and-mouth disease virus. a unique characteristic of these viruses is the use of a viral peptide (vpg) as primer for viral rna synthesis. as a consequence, all newly formed viral rna molecules possess a covalently linked vpg peptide. it is known that vpg is enzymatically released from the incoming viral rna by a host p ...201424243841
preclinical evaluation of the immunogenicity and safety of an inactivated enterovirus 71 candidate vaccine.human enterovirus 71 (ev71) is a significant cause of morbidity and mortality from hand, foot and mouth disease (hfmd) and neurological complications, particularly in young children in the asia-pacific region. there are no vaccines or antiviral therapies currently available for prevention or treatment of hfmd caused by ev71. therefore, the development of therapeutic and preventive strategies against hfmd is of growing importance. we report the immunogenic and safety profile of inactivated, purif ...201324244774
antiviral activity of guiqi polysaccharides against enterovirus 71 in vitro.in this study, we have investigated the antiviral activity of guiqi polysaccharides (gqp) upon enterovirus 71 (ev71) in vitro. an assay using methyl thiazolyl tetrazolium (mtt), and analyses of cytopathic effects (cpe) were used to examine the antiviral activity of gqp upon vero cells infected with ev71. the results revealed that gqp at concentrations below 31.2 μg/ml exhibited significant antiviral effects upon ev71 when applied under three different experimental protocols. gqp was most strongl ...201324254889
the cross-neutralizing activity of enterovirus 71 subgenotype c4 vaccines in healthy chinese infants and children.ev71 is one of major etiologic causes of hand-foot-mouth disease (hfmd) and leads to severe neurological complications in young children and infants. recently inactivated ev71 vaccines have been developed by five manufactures and clinically show good safety and immunogenicity. however, the cross-neutralizing activity of these vaccines remains unclear, and is of particular interest because rna recombination is seen more frequently in ev71 epidemics.201324260259
a 3c(pro)-dependent bioluminescence imaging assay for in vivo evaluation of anti-enterovirus 71 agents.enterovirus 71 (ev71), a member of picornaviridae, is one of the major pathogens of human hand, foot and mouth disease. ev71 mainly infects children and causes severe neurological complications and even death. the pathogenesis of ev71 infection is largely unknown, and no clinically approved vaccine or effective treatment is available to date. here we described a novel bioluminescence imaging approach for ev71 detection. in this approach, a plasmid-based reporter was constructed to express the fu ...201424263113
development and evaluation of a real-time method of simultaneous amplification and testing of enterovirus 71 incorporating a rna internal control system.human enterovirus 71 (ev71) is a primary etiological agent of hand, foot, and mouth disease (hfmd). the associated infections have posed a serious threat to the public health. in the present study, a real-time simultaneous amplification and testing (sat) technology was developed for detecting ev71 (sat-ev71). the rna of ev71 and an internal control (ic) were amplified and analyzed simultaneously by isothermal amplification and real-time detection of fluorescence using routine real-time pcr. furt ...201424269796
formalin-inactivated ev71 vaccine candidate induced cross-neutralizing antibody against subgenotypes b1, b4, b5 and c4a in adult volunteers.enterovirus 71 (ev71) has caused several epidemics of hand, foot and mouth diseases (hfmd) in asia. no effective ev71 vaccine is available. a randomized and open-label phase i clinical study registered with clinicaltrials.gov #nct01268787, aims to evaluate the safety, reactogenicity and immunogenicity of a formalin-inactivated ev71 vaccine candidate (ev71vac) at 5- and 10-µg doses. in this study we report the cross-neutralizing antibody responses from each volunteer against different subgenotype ...201324278177
how to understand the efficacy measurements for enterovirus type 71 vaccine?the choice of endpoint was most important for an efficacy vaccine trial. the objective of this paper is to gear toward answering questions about the rationality and scientificity of the primary endpoints choosing, case capturing and diagnosis strategy in our recently reported ev71 vaccine efficacy phase 3 trial. in order to obtain both high sensitivity and specificity in the case detecting, ev71-associated disease had been chosen as primary endpoint, a broad spectrum of clinical symptoms was sur ...201424280898
global impact of heparin on gene expression profiles in neural cells infected by enterovirus 71.heparan sulphate mimetics, particularly heparin (hep), have previously been shown to considerably inhibit infection of enterovirus 71 (ev71) in vero cells. therefore, in this study, a genome-wide dna microarray was performed to gain insight into the mechanism(s) of action of hep against infection of a human neural cell line, sk-n-sh, with a clinical strain of ev71.201424281657
a colorimetric-based accurate method for the determination of enterovirus 71 titer.the 50 % tissue culture infectious dose (tcid50) is still one of the most commonly used techniques for estimating virus titers. however, the traditional tcid50 assay is time consuming, susceptible to subjective errors and generates only quantal data. here, we describe a colorimetric-based approach for the titration of enterovirus 71 (ev71) using a modified method for making virus dilutions. in summary, the titration of ev71 using mtt or mts staining with a modified virus dilution method decrease ...201224293817
peripheral t lymphocyte subset imbalances in children with enterovirus 71-induced hand, foot and mouth disease.inflammatory mediators (i.e. cytokines) play a pivotal role in the regulation of pathophysiological processes during ev71-induced hand, foot and mouth disease (hfmd). different t cell subsets have distinct cytokine secretion profiles, and alteration in the t cell subsets frequency (imbalance) during infection leads to changed cytokine patterns. however, the effects of ev71 infection on t cell subsets were not clear. the objective of this study was to determine whether ev71-induced hfmd can be ex ...201424316007
the nuclear protein sam68 is redistributed to the cytoplasm and is involved in pi3k/akt activation during ev71 infection.nuclear proteins can be triggered to be redistributed to the cytoplasm to assist with ev71 virus replication. this process is frequently involved in cellular signal transduction upon virus infection. in this study, we have demonstrated that a new nuclear protein, 68-kda src-associated in mitosis protein (sam68), was translocated to the cytoplasm and was co-localized with ev71 during virus infection. confocal microscopy and subcellular fractionation assay confirmed that virus 3c protease triggere ...201424316008
[sero-epidemiological analysis for entervirus 71 infection of adults in beijing].to evaluate the enterovirus 71 (ev 71) protective antibody level of health adults people in beijing.201324319947
[development of ev71, ca16 and other enterovirus vrial real-time qualitative pcr diagnostic kit].a novel multiplex real-time rt-pcr kit was developed to detect ev71, coxa16 and other human enteroviruses simultaneously with an internal amplification control to avoids false negatives, which used for hand, foot and mouth disease in the clinical diagnosis and epidemiological surveillance.201324319965
immunization of n terminus of enterovirus 71 vp4 elicits cross-protective antibody responses.enterovirus 71 (ev71) is major cause of hand, foot and mouth disease. large epidemics of ev71 infection have been recently reported in the asian-pacific region. currently, no vaccine is available to prevent ev71 infection.201324320792
seroprevalence of human enterovirus 71 and coxsackievirus a16 in guangdong, china, in pre- and post-2010 hfmd epidemic period.human enterovirus 71 and coxsackie a16 have caused many outbreaks in the last decade in mainland china, resulting in thousands of fatal cases. seroepidemiology which provides important information to document population immunity is rare in china.201324324604
monoclonal antibodies for diagnosis of enterovirus 71.enterovirus 71 (ev71), one of the major causative agents of hand, foot, and mouth disease (hfmd), is now recognized as an emerging neurotropic virus in asia and may cause severe neurologic complications and mortalities. laboratory diagnosis of ev71 infection must be efficient and accurate, which could be accomplished by various immunoassays. in this study we use a live ev71 isolate, tainan/4643/98, with genotype c2 as an immunogen to sensitize balb/c (h-2(d)) mice and then generate the ev71-spec ...201324328741
dominant cd4-dependent rna-dependent rna polymerase-specific t-cell responses in children acutely infected with human enterovirus 71 and healthy adult controls.human enterovirus 71 (ev71) is one of the major causes of hand, foot and mouth disease (hfmd), which leads to significant mortality in infected children. a prophylactic vaccine is urgently needed. however, little is known about the protective t-cell immunity in individuals infected with the ev71 virus. in this study, we performed a comprehensive ex vivo interferon-γ elispot analysis in 31 children infected with ev71 as well as in 40 healthy adult controls of the cd4(+) and cd8(+) t-cell response ...201424329688
combined approaches to flexible fitting and assessment in virus capsids undergoing conformational change.fitting of atomic components into electron cryo-microscopy (cryoem) density maps is routinely used to understand the structure and function of macromolecular machines. many fitting methods have been developed, but a standard protocol for successful fitting and assessment of fitted models has yet to be agreed upon among the experts in the field. here, we created and tested a protocol that highlights important issues related to homology modelling, density map segmentation, rigid and flexible fitti ...201424333899
antigenic and genetic diversity of human enterovirus 71 from 2009 to 2012, taiwan.different subgenogroups of enterovirus 71 (ev-71) have caused numerous outbreaks of hand, foot, and mouth disease worldwide, especially in the asia-pacific region. during the development of a vaccine against ev-71, the genetic and antigenic diversities of ev-71 isolates from taiwan were analyzed by phylogenetic analyses and neutralization tests. the results showed that the dominant genogroups had changed twice, from b to c and from c to b, between 2009 and 2012. the subgenogroup b5 (b5b cluster) ...201324348916
human enterovirus 71 uncoating captured at atomic resolution.human enterovirus 71 (ev71) is the major causative agent of severe hand-foot-and-mouth diseases (hfmd) in young children, and structural characterization of ev71 during its life cycle can aid in the development of therapeutics against hfmd. here, we present the atomic structures of the full virion and an uncoating intermediate of a clinical ev71 c4 strain to illustrate the structural changes in the full virion that lead to the formation of the uncoating intermediate prepared for rna release. alt ...201424352461
the mechanism of translation initiation on type 1 picornavirus iress.picornavirus type 1 iress comprise five principal domains (dii-dvi). whereas dv binds eif4g, a conserved aug in dvi was suggested to stimulate attachment of 43s ribosomal preinitiation complexes, which then scan to the initiation codon. initiation on type 1 iress also requires ires trans-acting factors (itafs), and several candidates have been proposed. here, we report the in vitro reconstitution of initiation on three type 1 iress: poliovirus (pv), enterovirus 71 (ev71), and bovine enterovirus ...201424357634
construction of an infectious cdna clone of enterovirus 71: insights into the factors ensuring experimental success.enterovirus 71 (ev 71) is a causative agent of mild hand foot and mouth disease but is capable of causing severe complications in the cns in young children. reverse genetics technology is currently widely used to study the pathogenesis of the virus. the aim of this work was to determine and evaluate the factors which can contribute to infectivity of ev 71 rna transcripts in vitro. two strategies, overlapping rt-pcr and long distance rt-pcr, were employed to obtain the full-length genome cdna clo ...201424361875
complete genome sequence of a human enterovirus 71 strain isolated in wuhan, china, in 2010.the complete genome sequence of a human enterovirus 71 strain (ev71/wuhan/3018/2010), which was isolated in wuhan in 2010, was amplified by a reverse transcription-pcr method and sequenced. phylogenetic analysis based on the complete genome sequence classified this strain into subgenogroup a.201324371206
suramin inhibits ev71 infection.enterovirus-71 (ev71) is one of the major causative reagents for hand-foot-and-mouth disease. in particular, ev71 causes severe central nervous system infections and leads to numerous dead cases. although several inactivated whole-virus vaccines have entered in clinical trials, no antiviral agent has been provided for clinical therapy. in the present work, we screened our compound library and identified that suramin, which has been clinically used to treat variable diseases, could inhibit ev71 p ...201424374150
[analysis of pathogen spectrum of encephalitis/meningitis in northwestern area of china].to learn the characteristics of pathogen spectrum of encephalitis /meningitis in northwestern area of china.201324378133
tonsillar crypt epithelium is an important extra-central nervous system site for viral replication in ev71 encephalomyelitis.enterovirus 71 (ev71; family picornaviridae, species human enterovirus a) usually causes hand, foot, and mouth disease, which may rarely be complicated by fatal encephalomyelitis. we investigated extra-central nervous system (extra-cns) tissues capable of supporting ev71 infection and replication, and have correlated tissue infection with expression of putative viral entry receptors, scavenger receptor b2 (scarb2), and p-selectin glycoprotein ligand-1 (psgl-1). formalin-fixed, paraffin-embedded ...201424378407
enterovirus 2apro targets mda5 and mavs in infected cells.rig-i-like receptors (rlrs) mda5 and rig-i are key players in the innate antiviral response. upon recognition of viral rna, they interact with mavs, eventually inducing type i interferon production. the interferon induction pathway is commonly targeted by viruses. how enteroviruses suppress interferon production is incompletely understood. mda5 has been suggested to undergo caspase- and proteasome-mediated degradation during poliovirus infection. additionally, mavs is reported to be cleaved duri ...201424390337
molecular epidemiology of enterovirus 71 infection in the central region of taiwan from 2002 to 2012.enterovirus 71 (ev71), a causative agent of hand, foot, and mouth disease can be classified into three genotypes and many subtypes. the objectives of this study were to conduct a molecular epidemiological study of ev71 in the central region of taiwan from 2002-2012 and to test the hypothesis that whether the alternative appearance of different ev71 subtypes in taiwan is due to transmission from neighboring countries or from re-emergence of pre-existing local strains. we selected 174 ev71 isolate ...201324391812
the gene expression profile of peripheral blood mononuclear cells from ev71-infected rhesus infants and the significance in viral pathogenesis.enterovirus 71 (ev71) is the major pathogen responsible for fatal hand, foot and mouth disease (hfmd). our previous work reported on an ev71-infected rhesus monkey infant model that presented with histo-pathologic changes of the central nervous system (cns) and lungs. this study is focused on the correlated modulation of gene expression in the peripheral blood mononuclear cells (pbmcs) from ev71-infected rhesus monkey infants. the expression of more than 500 functional genes associated with mult ...201424392094
prevalence of multiple enteroviruses associated with hand, foot, and mouth disease in shijiazhuang city, hebei province, china: outbreaks of coxsackieviruses a10 and b3.hand, foot, and mouth disease (hfmd) has been one of the most common infectious diseases in shijiazhuang city, as is the situation in china overall. in the national hfmd surveillance system, the pathogen detection was focused on ev-a71 and cva16, and therefore, information on the other evs is very limited. in order to identify the circulating ev serotypes in the hfmd outbreaks in shijiazhuang city during 2010-2012, 4045 patients presented with hfmd were recruited in the study, and clinical sampl ...201424392117
spatio-temporal analysis on enterovirus cases through integrated surveillance in taiwan.severe epidemics of enterovirus have occurred frequently in malaysia, singapore, taiwan, cambodia, and china, involving cases of pulmonary edema, hemorrhage and encephalitis, and an effective vaccine has not been available. the specific aim of this study was to understand the epidemiological characteristics of mild and severe enterovirus cases through integrated surveillance data.201424400725
[observation on virus shedding periods of enterovirus-71 and coxsackievirus a 16 monitored by nucleic acids determination in stool samples of children with hand, foot and mouth disease].to observe the duration of enterovirus-71 (ev71) and coxsackievirus a 16 (coxa16) viral shedding in stool samples of children with hand, foot and mouth disease (hfmd) infected with ev71 and coxa16 and to explore the relationship between the duration of intestinal virus shedding and the severity of illness of children with hfmd.201324406235
interleukin-8 is elevated in severe hand, foot, and mouth disease.enterovirus 71 (ev71) infections can cause hand, foot, and mouth disease (hfmd), which is a potentially fatal illness in children. epidemics of hfmd are seen every year globally and present an increasing threat to public health worldwide.201424423718
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