• enterovirus 71
• life cycle
• virus–host interaction

• Humans
• Enterovirus A, Human/physiology
• Enterovirus A, Human/genetics
• Enterovirus A, Human/growth & development
• Virus Replication
• Enterovirus Infections/virology
• Viral Tropism
• Animals
• Host-Pathogen Interactions

• 32070184 and 32470144 the National Natural Science Foundation of China

• conjunctivitis
• disease outbreaks
• enterovirus

• Adult
• Humans
• Male
• Female
• Adolescent
• Young Adult
• Middle Aged
• Outpatients
• Tertiary Care Centers
• Cross-Sectional Studies
• Conjunctivitis/epidemiology
• Acute Disease

• Evolutionary game theory
• epidemic model
• imperfect vaccination

• Humans
• Commerce
• Game Theory
• Internationality
• Vaccination
• Epidemics/prevention & control

• Humans
• Conjunctivitis, Viral/diagnosis
• Conjunctivitis
• Adenoviridae Infections
• Adenoviridae
• Simplexvirus
• Keratitis, Herpetic
• Adenovirus Infections, Human/diagnosis
• Adenovirus Infections, Human/epidemiology
• Adenoviruses, Human

• CV-A6
• epidemiology
• hand, foot, and mouth disease clusters
• phylogenetic analysis

Honeysuckle (Lonicera japonica Thunb) is a traditional Chinese medicine (TCM) with an antipathogenic activity. MicroRNAs (miRNAs) are small non-coding RNA molecules that are ubiquitously expressed in cells. Endogenous miRNA may function as an innate response to block pathogen invasion. The miRNA expression profiles of both mice and humans after the ingestion of honeysuckle were obtained. Fifteen overexpressed miRNAs overlapped and were predicted to be capable of targeting three viruses: dengue virus (DENV), enterovirus 71 (EV71) and SARS-CoV-2. Among them, let-7a was examined to be capable of targeting the EV71 RNA genome by reporter assay and Western blotting. Moreover, honeysuckle-induced let-7a suppression of EV71 RNA and protein expression as well as viral replication were investigated both in vitro and in vivo. We demonstrated that let-7a targeted EV71 at the predicted sequences using luciferase reporter plasmids as well as two infectious replicons (pMP4-y-5 and pTOPO-4643). The suppression of EV71 replication and viral load was demonstrated in two cell lines by luciferase activity, RT-PCR, real-time PCR, Western blotting and plaque assay. Furthermore, EV71-infected suckling mice fed honeysuckle extract or inoculated with let-7a showed decreased clinical scores and a prolonged survival time accompanied with decreased viral RNA, protein expression and virus titer. The ingestion of honeysuckle attenuates EV71 replication and related pathogenesis partially through the upregulation of let-7a expression both in vitro and in vivo. Our previous report and the current findings imply that both honeysuckle and upregulated let-7a can execute a suppressive function against the replication of DENV and EV71. Taken together, this evidence indicates that honeysuckle can induce the expression of let-7a and that this miRNA as well as 11 other miRNAs have great potential to prevent and suppress EV71 replication.

• EV71
• honeysuckle
• let-7a
• virus replication

• Animals
• Antiviral Agents/pharmacology
• Cell Line
• Enterovirus A, Human/drug effects
• Enterovirus A, Human/physiology
• Enterovirus Infections/drug therapy
• Humans
• Lonicera/chemistry
• Mice
• Mice, Inbred ICR
• MicroRNAs/metabolism
• Plant Extracts/pharmacology
• Virus Replication/drug effects

During recent 20 years, enterovirus 71 (EV71) has emerged as a major concern among children, particularly in the Asia-Pacific region. To understand current EV71 serostatus, to find risk factors associated with EV71 infection and to establish future EV71 vaccine policy, we performed a seroepidemiology study in Taiwan in 2017.

After informed consent was obtained, we enrolled preschool children, 6–15-year-old students, 16–50-year-old people. They received a questionnaire and a blood sample was collected to measure the EV71 neutralization antibody.

Altogether, 920 subjects were enrolled with a male-to-female ratio of 1.03. The EV71 seropositive rate was 10% (8/82) in infants, 4% (6/153) in 1-year-old children, 8% (7/83) in 2-year-old children, 8% (13/156) in 3–5-year-old children, 31% (38/122) in 6–11-year-old primary school students, 45% (54/121) in 12–15-year-old high school students and 75% (152/203) in 16-50-year-old people. Risk factors associated with EV71 seropositivity in preschool children were female gender, having siblings, more siblings, and contact with herpangina or hand-foot-and-mouth disease. The risk factor with EV71 seropositivity in 16–50-year-old people was having children in their families in addition to older age (p<0.001). Compared with the rates in 1997, 1999 and 2007, the rates in children were significantly lower in 2017.

EV71 seropositive rates were very low, at 4% to 10%, in preschool children and not high, at 31%, in primary school students. Preschool children are highly susceptible and need EV71 vaccine most.

• Adolescent
• Adult
• Antibodies, Neutralizing/blood
• Child
• Child, Preschool
• Enterovirus A, Human/immunology
• Enterovirus A, Human/isolation & purification
• Enterovirus Infections/complications
• Enterovirus Infections/diagnosis
• Enterovirus Infections/epidemiology
• Enterovirus Infections/virology
• Female
• Hand, Foot and Mouth Disease/complications
• Hand, Foot and Mouth Disease/diagnosis
• Herpangina/complications
• Herpangina/diagnosis
• Humans
• Infant
• Male
• Middle Aged
• Risk Factors
• Sex Factors
• Taiwan/epidemiology
• Young Adult

• The Taiwan Centers for Disease Control, the Ministry of Health and Welfare
• the Ministry of Science and Technology, Taiwan

• brain stem/cerebellum
• haematology (incl blood transfusion)
• infection (neurology)
• neuro Itu

• Biochemistry
• Biological Sciences
• Cell Biology
• Microbiology
• Molecular Microbiology
• Viral Microbiology
• Virology

• Age of onset
• Attention deficiency hyperactivity disorder
• Cognition
• Encephalitis
• Enterovirus A71
• Polio-like syndrome
• Rhombencephalitis
• Sequelae
• Tracheostomy

• Complexity
• Diversity
• Mild HFMD
• Next-generation sequencing
• Severe HFMD
• Virome

Enterovirus 71 (EV71) is the predominant causative pathogen of hand-foot-and-mouth disease (HFMD). Contrary to other HFMD-causing enterovirus, EV71 can lead to severe neurological complications, even death. MicroRNAs (miRNAs) are small non-coding RNAs that constitute the largest family of gene regulators participating in numerous biological or pathological processes. We previously reported that miR-16-5p increases with severity of HFMD by investigating the expression patterns of host miRNAs in patients with HFMD. However, the mechanisms by which EV71 induces miR-16-5p expression are not clear, and the interaction between EV71 and miR-16-5p is not yet fully understood. Here, we confirmed EV71-induced expression of miR-16-5p both in vitro and in vivo and show that upregulation of miR-16-5p by EV71 infection may occur at the posttranscriptional level. Moreover, EV71-induced caspase activation facilitates the processing of pri-miR-16-1. We also revealed that miR-16-5p can promote EV71-induced nerve cells apoptosis through activating caspase-3. In addition, we found that miR-16-5p can inhibit EV71 replication. CCNE1 and CCND1, two important cell cycle regulators, play an important role in the suppression of EV71 replication by miR-16-5p. Therefore, miR-16-5p is a positive feedback regulator in EV71-induced apoptosis and a suppressor of virus replication. These results help in understanding the interaction network between miRNA and EV71 infection and provide a potential target for the development of antiviral therapy.

Enterovirus A71 (EV-A71) causes hand-foot-and-mouth disease (HFMD), which may be complicated by fatal encephalomyelitis. Although fecal–oral or oral–oral routes are important in person-to-person transmission, how viral shedding and exposure may predispose individuals to infection remains unknown. We investigated person-to-person transmission by using a model of HFMD and encephalomyelitis based on EV-A71 oral infection of 2-week-old hamsters. Animals (index animals) infected with 10⁴ 50% cell culture infective doses of virus uniformly developed severe disease four days post-infection (dpi), whereas littermate contacts developed severe disease after six to seven days of exposure to index animals. Virus was detected in oral washes and feces at 3–4 dpi in index animals and at three to eight days after exposure to index animals in littermate contact animals. In a second experiment, non-littermate contact animals exposed for 8 or 12 h to index animals developed the disease six and four days post-exposure, respectively. Tissues from killed index and contact animals, studied by light microscopy, immunohistochemistry and in situ hybridization, exhibited mild inflammatory lesions and/or viral antigens/RNA in the squamous epithelia of the oral cavity, tongue, paws, skin, esophagus, gastric epithelium, salivary glands, lacrimal glands, central nervous system neurons, muscles (skeletal, cardiac and smooth muscles) and liver. Orally shed viruses were probably derived from infected oral mucosa and salivary glands, whereas fecal viruses may have derived from these sites as well as from esophageal and gastric epithelia. Asymptomatic seroconversion in exposed mother hamsters was demonstrated. Our hamster model should be useful in studying person-to-person EV-A71 transmission and how drugs and vaccines may interrupt transmission.

• Case-control study
• China
• Hand, foot, and mouth disease
• Handwashing
• Transmission

• Case-Control Studies
• Child, Preschool
• China/epidemiology
• Enterovirus/physiology
• Feces/virology
• Female
• Hand Disinfection
• Hand, Foot and Mouth Disease/epidemiology
• Hand, Foot and Mouth Disease/transmission
• Hand, Foot and Mouth Disease/virology
• Humans
• Infant
• Infection Control
• Male
• Mouth/virology

• Contaminated environment
• HFMD
• Indirect transmission
• Uniform persistence

• Causality
• Child
• Child, Preschool
• China/epidemiology
• Computer Simulation
• Environmental Exposure/statistics & numerical data
• Environmental Pollution/statistics & numerical data
• Female
• Hand, Foot and Mouth Disease/mortality
• Hand, Foot and Mouth Disease/transmission
• Hand, Foot and Mouth Disease/virology
• Humans
• Infant
• Infant, Newborn
• Male
• Models, Statistical
• Prevalence
• Reproducibility of Results
• Risk Assessment/methods
• Risk Factors
• Sensitivity and Specificity
• Survival Analysis

Epidemics of hand, foot and mouth disease (HFMD) among children in East Asia have been a serious annual public health problem. Previous studies in China and island-type territories in East Asia showed that the onset of HFMD epidemics evolved with increased latitude. Based on the natural characteristics of the epidemics, we developed regression models for issuing aberration alerts and predictions.

HFMD sentinel surveillance data from 2008 to 2014 in Japan are used in this study, covering 365 weeks and 47 prefectures between 24 and 46° of north latitude. Average HFMD cases per sentinel are standardized as Z rates. We fit weekly Z rate differences between prefectures located in the south and north of a designated prefecture with linear regression models to detect the surging trend of the epidemic for the prefecture. We propose a rule for issuing an aberration alert determined by the strength of the upward trend of south–north Z rate differences in the previous few weeks. In addition to the warning, we predict a Z rate for the next week with a 95 % confidence interval.

We selected Tokyo and Kyoto for evaluating the proposed approach to aberration detection. Overall, the peaks of epidemics in Tokyo mostly occurred in weeks 28–31, later than in Kyoto, where the disease peaked in weeks 26–31. Positive south–north Z rate differences in both prefectures were clearly observed ahead of the HFMD epidemic cycles. Aberrations in the major epidemics of 2011 and 2013 were successfully detected weeks earlier. The prediction also provided accurate estimates of the epidemic’s trends.

We have used only the latitude, one geographical feature affecting the spatiotemporal distribution of HFMD, to develop rules for early aberration detection and prediction. We have also demonstrated that the proposed rules performed well using real data in terms of accuracy and timeliness. Although our approach may provide helpful information for controlling epidemics and minimizing the impact of diseases, the performance could be further improved by including other influential meteorological factors in the proposed latitude-based approach, which is worth further investigation.

Korean Red Ginseng (KRG) is a heat-processed ginseng developed by the repeated steaming and air-drying of fresh ginseng. Compared with fresh ginseng, KRG has been shown to possess greater pharmacological activities and stability because of changes that occur in its chemical constituents during the steaming process. In addition to anticancer, anti-inflammatory, and immune-modulatory activities, KRG and its purified components have also been shown to possess protective effects against microbial infections. Here, we summarize the current knowledge on the properties of KRG and its components on infections with human pathogenic viruses such as respiratory syncytial virus, rhinovirus, influenza virus, human immunodeficiency virus, human herpes virus, hepatitis virus, norovirus, rotavirus, enterovirus, and coxsackievirus. Additionally, the therapeutic potential of KRG as an antiviral and vaccine adjuvant is discussed.

• Korean Red Ginseng
• antiviral
• ginsenosides
• hepatitis virus
• influenza virus

Enterovirus A71 (EVA71) and Coxsackievirus A16 (CVA16) are regarded as the two major causative pathogens in hand, foot and mouth disease (HFMD) epidemics. However, CVA6, previously largely ignored, became the predominant pathogen in China in 2013. In this study, we describe the epidemiological trendsofCVA6 during the annual HFMD outbreaks from 2008 to 2013 in Guangdong, China. The study results show that CVA6 has been one of three major causative agents of HFMD epidemics since 2009. The periodic rotation and dominance of the three pathogens, EVA71, CVA16 and CVA6, may have contributed to the continuously increasing HFMD epidemics. Moreover, phylogenetic analysis of the VP1 gene shows that major circulating CVA6 strains collected from 2009 to 2013 are distinct from the earlier strains collected before 2009. In conclusion, the discovery from this research investigating epidemiological trends of CVA6 from 2008 to 2013 explains the possible pattern of the continuous HFMD epidemic in China. The etiological change pattern also highlights the need for improvement for pathogen surveillance and vaccine strategies for HFMD control in China.

The person-to-person transmission of the hepatitis A virus primarily occurs in enclosed spaces, particularly in the presence of inadequate hygiene conditions and a high proportion of susceptible individuals. Thus, intimate family contact stands out as a risk factor for HAV infection dissemination. The present study aimed to evaluate the occurrence of household HAV transmission. Blood samples were collected from patients with hepatitis A (index cases) and their family members (contacts) that were referred to an ambulatory care clinic specializing in viral hepatitis. A total of 97 samples were collected from 30 families with a confirmed hepatitis A case (index case). Serological and molecular techniques for the diagnosis of hepatitis A were conducted on all samples. HAV infection (anti-HAV IgM + and/or HAV RNA +) was detected in 34.3% (23/67) of the contacts; 34.3% (23/67) of the contacts were immune to HAV, and 31.4% (21/67) were susceptible. In the household contacts, HAV immunity was significantly associated with older age; susceptibility to infection and HAV infection were associated with younger age. Household outbreaks were detected in 16/30 families studied. Co-circulation of subgenotypes IA and IB was found in the household outbreaks, and person-to-person transmission was evidenced in six of the household outbreaks, with 100% homology between the index case and contact strains. The results demonstrated the relevance of HAV household transmission, reaffirming the need for hepatitis A vaccine administration in susceptible contacts and effective infection control procedures to prevent the extension of household outbreaks.

The VP1 protein of enterovirus 71 (EV71) is an important immunodominant protein which is responsible for host-receptor binding. Nevertheless, the relationship between VP1 and neurovirulence is still poorly understood. In this study, we investigated the relationship between mutation of VP1 and neurovirulent phenotype of EV71 infection.

One hundred and eighty-seven strains from Genbank were included, with a clear clinical background. They were divided into two groups, one with nervous system symptoms and one with no nervous system symptoms. After alignment, the significance of amino acid variation was determined by using the χ² test and a phylogenetic tree was constructed with MEGA software (version 5.1).

We showed no significant difference in neurovirulence between genotype B and C. Interestingly, we found that variations of E145G/Q, E164D/K and T292N/K were associated with nervous system infection in genotype B. In the case of genotype C, the N31D mutation increased the risk for nervous complications, whereas I262V mutation decreased the risk of nervous complications. We used a 3D model of VP1 to demonstrate the potential molecular basis for EV71 nervous system tropism.

Distinct variations are shown to be associated with neurovirulent phenotype in the different genotype. Detection of variation in genotypes and subtypes may be important for the prediction of clinical outcomes.

Hand, foot and mouth disease (HFMD) is an important public health problem that has emerged over the past several years. HFMD predominantly infects children under seven years old and occasionally causes severe disease in adults. Among the enteroviruses, enterovirus 71 (EV71) and coxsackievirus 16 (CA16) are the major causative agents of HFMD. In addition, adenovirus cocirculates with enterovirus and has become a possible additional pathogenic factor for HFMD in some cases. Here, we have investigated the neutralizing antibody responses to both enterovirus and adenovirus in adults, with the aim of exploring the prevalence trends of these viruses and the nature of protective immunity in humans to these viral infections. Sera from 391 healthy adults from 21 provinces and cities in China were tested for the presence of antibodies against EV71, CA16, adenovirus human serotype 5 (AdHu5) and chimpanzee adenovirus pan7 (AdC7) using neutralization tests. High seroprevalence rates of EV71, CA16 and AdHu5 were found in the population (85.7%, 58.8% and 74.2%, respectively). The coseropositivity rate of these three viruses was 39.4% (154 of 391), with median neutralizing antibody titers of 80, 40 and 640, respectively, and the neutralizing antibody titer for EV71 was found to be correlated with those of CA16 and AdHu5. AdC7 was found to be a rare adenovirus serotype in the human population, with a seropositivity rate of 11.8%, suggesting that it could be a good choice for a vaccine carrier that could be used in vaccine development.

• AdC7
• AdHu5
• CA16
• EV71
• hand, foot and mouth disease
• neutralizing antibody
• seroprevalence

• Animals
• Cell Line, Tumor
• Child, Preschool
• China
• Chlorocebus aethiops
• Enterovirus C, Human/genetics
• Enterovirus C, Human/isolation & purification
• Enterovirus C, Human/physiology
• Enterovirus Infections/virology
• Female
• Genome, Viral
• Humans
• Kinetics
• Paraplegia/virology
• Phylogeny
• Recombination, Genetic
• Sequence Analysis, DNA
• Vero Cells
• Viral Structural Proteins/genetics
• Virus Replication

• PCR
• adolescent
• autoantibodies
• biomarker
• child
• enterovirus
• family
• parent
• pathogenesis
• sibling
• type 1 diabetes
• virology
• virus

• Antiviral
• host-virus interaction
• human enterovirus
• interferon respons
• viral replication

Outbreaks of hand, foot, and mouth disease caused by enterovirus 71 (EV71) have become considerable threats to the health of infants and young children. To identify the cellular long noncoding RNAs (lncRNAs) involved in the host response to EV71 infection, we performed comprehensive lncRNA and mRNA profiling in EV71-infected rhabdomyosarcoma cells through microarray. We observed the differential expression of more than 4800 lncRNAs during infection. Further analysis showed 160 regulated enhancer-like lncRNA and nearby mRNA pairs, as well as 313 regulated Rinn’s lncRNA [M. Guttman I. Amit, M. Garber, C. French, M.F. Lin, D. Feldser, M. Huarte, O. Zuk, B.W. Carey, J.P. Cassady, M.N. Cabili, R. Jaenisch, T.S. Mikkelsen, T. Jacks, N. Hacohen, B.E. Bernstein, M. Kellis, A. Regev, J.L. Rinn, E.S. Lander. Chromatin signature reveals over a thousand highly conserved large non-coding RNAs in mammals. Nature 458 (2009) 223–227, A.M. Khalil, M. Guttman, M. Huarte, M. Garber, A. Raj, D. Rivea Morales, K. Thomas, A. Presser, B.E. Bernstein, A. van Oudenaarden, A. Regev, E.S. Lander, J.L. Rinn. Many human large intergenic noncoding RNAs associate with chromatin-modifying complexes and affect gene expression. Proc. Natl. Acad. Sci. USA 106 (2009) 11667–11672] and nearby mRNA pairs. The results provided information for further research on the prevention and treatment of EV71 infection, as well as on distinguishing severe and mild EV71 cases.

• hand-foot-mouth disease
• enterovirus 71
• co-detection

• Bocavirus/isolation & purification
• Child, Preschool
• Coinfection/pathology
• Coinfection/virology
• Enterovirus A, Human/isolation & purification
• Feces/virology
• Female
• Hand, Foot and Mouth Disease/pathology
• Hand, Foot and Mouth Disease/virology
• Humans
• Infant
• Male
• Norovirus/isolation & purification
• Rotavirus/isolation & purification

• enterovirus 71
• macrophages
• proinflammatory cytokines
• chemokines
• toll-like receptors

• Adult
• Cells, Cultured
• Cytokines/metabolism
• DEAD Box Protein 58
• DEAD-box RNA Helicases/biosynthesis
• Enterovirus A, Human/immunology
• Gene Expression Profiling
• Humans
• Interferon-Induced Helicase, IFIH1
• Macrophages/immunology
• Macrophages/virology
• Real-Time Polymerase Chain Reaction
• Receptors, Immunologic
• Toll-Like Receptors/biosynthesis
• Young Adult

• Disinfectants/pharmacology
• Enterovirus/drug effects
• Enterovirus A, Human/drug effects
• Hand, Foot and Mouth Disease/prevention & control
• Humans
• Microbial Sensitivity Tests
• Sodium Hypochlorite/pharmacology
• Time Factors
• Virus Inactivation

• Age Factors
• Child
• Child, Preschool
• China/epidemiology
• Delayed Diagnosis/statistics & numerical data
• Disease Outbreaks/statistics & numerical data
• Enterovirus A, Human/pathogenicity
• Female
• Hand, Foot and Mouth Disease/epidemiology
• Hand, Foot and Mouth Disease/transmission
• Hand, Foot and Mouth Disease/virology
• Humans
• Infant
• Male
• Population Surveillance
• Risk Factors
• Seasons
• Severity of Illness Index
• Sex Factors

• R01 CA095994 NCI NIH HHS
• R01 CA095994-04 NCI NIH HHS
• U01-GM070749 NIGMS NIH HHS
• U01 GM070749 NIGMS NIH HHS
• U01 GM070749-10 NIGMS NIH HHS

AIM: To characterise the viral kinetics of enterovirus 71 (EV71).

METHODS: In this study, human rhabdomyosarcoma (RD) cells were infected with EV71 at different multiplicity of infection (MOI). After infection, the cytopathic effect (CPE) was monitored and recorded using a phase contrast microscope associated with a CCD camera at different time points post viral infection (0, 6, 12, 24 h post infection). Cell growth and viability were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in both EV71 infected and mock infected cells at each time point. EV71 replication kinetics in RD cells was determined by measuring the total intracellular viral RNA with real-time reverse-transcription polymerase chain reaction (qRT-PCR). Also, the intracellular and extracellular virion RNA was isolated and quantified at different time points to analyze the viral package and secretion. The expression of viral protein was determined by analyze the levels of viral structure protein VP1 with Western blotting.

RESULTS: EV71 infection induced a significant CPE as early as 6 h post infection (p.i.) in both RD cells infected with high ratio of virus (MOI 10) and low ratio of virus (MOI 1). In EV71 infected cells, the cell growth was inhibited and the number of viable cells was rapidly decreased in the later phase of infection. EV71 virions were uncoated immediately after entry. The intracellular viral RNA began to increase at as early as 3 h p.i. and the exponential increase was found between 3 h to 6 h p.i. in both infected groups. For viral structure protein synthesis, results from western-blot showed that intracellular viral protein VP1 could not be detected until 6 h p.i. in the cells infected at either MOI 1 or MOI 10; and reached the peak at 9 h p.i. in the cells infected with EV71 at both MOI 1 and MOI 10. Simultaneously, the viral package and secretion were also actively processed as the virus underwent rapid replication. The viral package kinetics was comparable for both MOI 1 and MOI 10 infected groups. It was observed that at 3 h p.i, the intracellular virions obviously decreased, thereafter, the intracellular virions began to increase and enter into the exponential phase until 12 h p.i. The total amounts of intracellular virons were decreased from 12 to 24 h p.i. Consistent with this result, the increase of virus secretion occurred during 6 to 12 h p.i.

CONCLUSION: The viral kinetics of EV71 were established by analyzing viral replication, package and secretion in RD cells.

• Enterovirus 71
• Quantitative reverse transcription polymerase chain reaction
• Viral kinetics
• Western blotting

Enterovirus 71 (EV71) has emerged as a significant pathogen causing large outbreaks in China for the past 3 years. Developing an EV71 vaccine is urgently needed to stop the spread of the disease; however, the adaptive immune response of humans to EV71 infection remains unclear. We examined the neutralizing antibody titers in HFMD patients and compared them to those of asymptomatic healthy children and young adults. We found that 80% of HFMD patients became positive for neutralizing antibodies against EV71 (GMT = 24.3) one day after the onset of illness. The antibody titers in the patients peaked two days (GMT = 79.5) after the illness appeared and were comparable to the level of adults (GMT = 45.2). Noticeably, the antibody response was not correlated with disease severity, suggesting that cellular immune response, besides neutralizing antibodies, could play critical role in controlling the outcome of EV71 infection in humans.

EV71 is associated with the fatal cases of brain stem encephalitis during large HFMD outbreaks from 1998 to 2008. EV71 may continuously shed from upper respiratory tracts and feces of HFMD patients for relatively long time after recovery. However, the persistence of viruses in the patients' secretions and excretions is not clear.

Serial throat swabs and feces of 34 definitely diagnosed patients, including 30 mild cases and 4 severe cases, were traced and collected with the interval of 2 to 4 days for up to 32 and 48 days, respectively, and tested by a nested RT-PCR.

The EV-71 specific sequences were identified by a Nested RT-PCR in all specimens of 0-4 days, and 5-8 days. The positive rates of EV71 in throat swabs dropped markedly to 42.86% during 9-12 days, and maintained at 20-30% during 13-24 days, while that in feces reduced to 71.43% during 9-12 days, and maintained roughly 20% till 37-40 days. EV71 nucleotide of 36.36% cases disappeared simultaneously both in throats and feces, 39.39% cases showed longer persistence of EV71 nucleotides in feces, and 21.21% were longer in throats. The longest duration of shedding observed was 24 days for throat swabs and 42 days for fecal specimens.

EV71 shedding from respiratory tract may continue for nearly four weeks after onset, but its excretion through feces can persist more than five weeks.

• Adolescent
• Adult
• Aged
• Aged, 80 and over
• Antibodies, Neutralizing/blood
• Antibodies, Viral/blood
• Child
• Child, Preschool
• Enterovirus/immunology
• Enterovirus A, Human/immunology
• Female
• Germany/epidemiology
• Hand, Foot and Mouth Disease/epidemiology
• Humans
• Infant
• Male
• Middle Aged
• Seroepidemiologic Studies
• Young Adult