• Coxsackievirus A6
• cross-neutralization
• cross-protection
• hand foot and mouth disease
• vaccine

• Adult
• Humans
• Animals
• Mice
• Rats
• Antibodies, Neutralizing
• Antibodies, Viral
• Vaccines
• Research
• China

• Hand, Foot and Mouth Disease/epidemiology
• Hand, Foot and Mouth Disease/virology
• Humans
• Enterovirus A, Human/genetics
• Enterovirus A, Human/pathogenicity
• Enterovirus/classification
• Enterovirus/genetics
• China/epidemiology

• CV-A6
• China
• HFMD
• coxsackievirus A6
• evolution
• hand, foot, and mouth disease
• molecular epidemiology

• Hand, Foot and Mouth Disease/epidemiology
• Hand, Foot and Mouth Disease/virology
• China/epidemiology
• Humans
• Evolution, Molecular
• Molecular Epidemiology
• Enterovirus A, Human/genetics
• Enterovirus A, Human/isolation & purification
• Enterovirus A, Human/classification
• Phylogeny
• Enterovirus/genetics
• Enterovirus/classification
• Enterovirus/isolation & purification
• Genomics
• Male

• viral epidemiology
• viral evolution
• retrovirus

• Humans
• Phylogeny
• Genotype
• China/epidemiology
• Enterovirus B, Human/genetics
• Recombination, Genetic
• Hand, Foot and Mouth Disease/epidemiology

• 2022YFC2305302 the National Key Research and Development Program of China

• molecular biology
• diseases
• molecular medicine

• Coxsackievirus A6
• Enterovirus
• Genetic variations
• HFMD
• Molecular epidemiology

Echovirus, a member of the Enterovirus B (EV-B) family, has led to numerous outbreaks and pandemics, causing a broad spectrum of diseases. Based on the national hand, foot, and mouth disease (HFMD) surveillance system, seven strains of echovirus 33 (E33) were isolated from Mainland of China between 2010 and 2018. The whole genomes of these strains were isolated and sequenced, and phylogenetic trees were constructed based on the gene sequences in different regions of the EV-B prototype strains. It was found that E33 may be recombined in the P2 and P3 regions. Five genotypes (A–E) were defined based on the entire VP1 region of E33, of which the C gene subtype was the dominant gene subtype at present. Recombinant analysis showed that genotype C strains likely recombined with EV-B80, EV-B85, E13, and CVA9 in the P2 and P3 regions, while genotype E had the possibility of recombination with CVB3, E3, E6, and E4. Results of Bayesian analysis indicated that E33 may have appeared around 1955 (95% confidence interval: 1945–1959), with a high evolutionary rate of 1.11 × 10⁻² substitution/site/year (95% highest posterior density (HPD): 8.17 × 10⁻³ to 1.4 × 10⁻² substitution/site/year). According to spatial transmission route analysis, two significant transmission routes were identified: from Australia to India and from Oman to Thailand, which the E33 strain in Mainland of China likely introduced from Mexico and India. In conclusion, our study fills the gaps in the evolutionary analysis of E33 and can provide important data for enterovirus surveillance.

Hand, foot, and mouth disease (HFMD) has become one of the most common infectious diseases in China. Before 2016, the primary causal serotypes were enterovirus A71 (EV-A71) and coxsackievirus A16 (CV-A16). Following the introduction of EV-A71 vaccines in China since 2016, the situation could change. CV-A6 has recently replaced EV-A71 and CV-A16 in some areas of China. However, the epidemiological characteristics of central China remain unknown.

To investigate the clinical symptoms and pathogen spectrum of HFMD in Shiyan City, central China, in recent years.

The epidemiological, clinical, and laboratory data from HFMD cases reported to the Shiyan Center for Disease Control and Prevention between January 2016 and December 2020 were analyzed. 196 throat swab specimens were collected from hospitalized HFMD patients between January 2018 and December 2020. To detect and genotype enteroviruses, real-time reverse transcription-polymerase chain reaction and sequencing of the 5'-untranslated region were used. In Shiyan, 168 laboratory-confirmed HFMD cases were studied using a logistic regression model to determine the effect of predominant enterovirus serotypes. Based on the logistic regression model, the least absolute shrinkage and selection operator model was used to analyze the correlation between CV-A6 infection and various clinical characteristics in HFMD patients in Shiyan.

From 2016 to 2020, 35840 HFMD cases were reported in Shiyan. The number of cases decreased by 48.4% from 2016 to 2017. Approximately 1.58-fold increases were found in 2018 and 2019 when compared to the previous year, respectively. In 2020, a decrease of about 85.5% was reported when compared to 2019. The most common serotypes shifted from EV-A71 and CV-A16 (about 60%-80% in 2016 and 2018) to others (more than 80.0% in 2017, 2019, and 2020). EV-A71 lost its dominance in 2017 in Shiyan. Among 196 confirmed HFMD cases, 85.7% tested positive for enterovirus, with CV-A6 being the most common serotype (121/168, 72.0%). The positive rates for CV-A16 and CV-A10 were 4.8% and 3.0%, respectively. There was no EV-A71 discovered. Infection with CV-A6 was linked to fever, myocardial damage, increased creatine kinase MB isoenzyme, and lactate dehydrogenase levels.

CV-A6 was the most common enterovirus serotype in Shiyan City, replacing EV-A71 and CV-A16 as the HFMD pathogen. Developing vaccines against CV-A6 or multiple pathogens, as well as rising CV-A6 surveillance, will help prevent HFMD in central China.

• Hand, foot, and mouth disease
• Coxsackievirus A6
• Enterovirus
• Prevalence serotype
• Fever
• Myocardial damage

After the first national-scale outbreak of Hand, foot, and mouth disease (HFMD) in China, a national surveillance network was established. Here we described the epidemiology and pathogenic profile of HFMD and the impact of EV-A71 vaccination on pathogen spectrum of enteroviruses in the southeastern Chinese city of Nanchang during 2010–2019. A total of 7,951 HFMD cases from sentinel hospitals were included, of which 4,800 EV-positive cases (60.4%) were identified by real-time RT-PCR. During 2010–2012, enterovirus 71 (EV-A71) was the main causative agent of HFMD, causing 63.1% of cases, followed by 19.3% cases associated with coxsackievirus A16 (CV-A16). Since 2013, the proportion of other enteroviruses has increased dramatically, with the sub genotype D3 strain of Coxsackievirus A6 (CV-A6) replacing the dominance of EV-A71. These genetically diverse native strains of CV-A6 have co-transmitted and co-evolved in Nanchang. Unlike EV-A71 and CV-A16, most CV-A6 infections were concentrated in autumn and winter. The incidence of EV-A71 infection negatively correlated with EV-A71 vaccination (r = −0.990, p = 0.01). And severe cases sharply declined as the promotion of EV-A71 vaccines. After 2-year implementation of EV-A71 vaccination, EV-A71 is no longer detected from the reported HFMD cases in Nanchang. In conclusion, EV-A71 vaccination changed the pattern of HFMD epidemic, and CV-A6 replaced the dominance of EV-A71 over time.

• Coxsackievirus A6
• EV-A71 vaccine
• HFMD
• epidemiology
• phylogenetic analysis
• surveillance

• Changing epidemiology
• Comparative study
• Enterovirus 71 vaccine
• HFMD

• clinical features
• enterovirus types
• hand foot and mouth disease
• herpangina
• molecular epidemiology

Hand, foot, and mouth disease (HFMD) is a common infectious disease affecting mainly children under 5 years of age. Coxsackievirus A6 (CVA-6), a major causative pathogen of HFMD, has caused outbreaks in recent years. Currently, no effective vaccine or antiviral treatments are available. In this study, one-step reverse-transcription recombinase polymerase amplification (RT-RPA), combined with a disposable lateral flow strip (LFS) assay, was developed to detect CVA-6. This assay can be performed in less than 35 min at 37°C without expensive instruments, and the result can be observed directly with the naked eye. The sensitivity of the RT-RPA-LFS was 10 copies per reaction, which was comparable to that of the conventional real-time quantitative polymerase chain reaction (qPCR) assays. Moreover, the assay specificity was 100%. The clinical performance of the RT-RPA-LFS assay was evaluated using 142 clinical samples, and the coincidence rate between RT-RPA-LFS and qPCR was 100%. Therefore, our RT-RPA-LFS assay provides a simple and rapid approach for point-of-care CVA-6 diagnosis.

• Coxsackievirus A6
• enterovirus
• lateral flow strips
• rapid detection
• recombinase polymerase amplification

• Hand
• Case-control study
• Coxsackievirus A6
• Hand-washing
• Risk factors
• foot and mouth disease

Hand, foot, and mouth disease (HFMD) is a common global epidemic. From 2008 onwards, many HFMD outbreaks caused by coxsackievirus A6 (CV-A6) have been reported worldwide. Since 2013, with a dramatically increasing number of CV-A6-related HFMD cases, CV-A6 has become the predominant HFMD pathogen in mainland China. Phylogenetic analysis based on the VP1 capsid gene revealed that subtype D3 dominated the CV-A6 outbreaks. Here, we performed a large-scale (near) full-length genetic analysis of global and Chinese CV-A6 variants, including 158 newly sequenced samples collected extensively in mainland China between 2010 and 2018. During the global transmission of subtype D3 of CV-A6, the noncapsid gene continued recombining, giving rise to a series of viable recombinant hybrids designated evolutionary lineages, and each lineage displayed internal consistency in both genetic and epidemiological features. The emergence of lineage –A since 2005 has triggered CV-A6 outbreaks worldwide, with a rate of evolution estimated at 4.17 × 10⁻³ substitutions site^-1 year⁻¹ based on a large number of monophyletic open reading frame sequences, and created a series of lineages chronologically through varied noncapsid recombination events. In mainland China, lineage –A has generated another two novel widespread lineages (–J and –L) through recombination within the enterovirus A gene pool, with robust estimates of occurrence time. Lineage –A, –J, and –L infections presented dissimilar clinical manifestations, indicating that the conservation of the CV-A6 capsid gene resulted in high transmissibility, but the lineage-specific noncapsid gene might influence pathogenicity. Potentially important amino acid substitutions were further predicted among CV-A6 variants. The evolutionary phenomenon of noncapsid polymorphism within the same subtype observed in CV-A6 was uncommon in other leading HFMD pathogens; such frequent recombination happened in fast-spreading CV-A6, indicating that the recovery of deleterious genomes may still be ongoing within CV-A6 quasispecies. CV-A6-related HFMD outbreaks have caused a significant public health burden and pose a great threat to children’s health; therefore, further surveillance is greatly needed to understand the full genetic diversity of CV-A6 in mainland China.

• Coxsackievirus A6
• evolutionary dynamics
• foot and mouth disease
• genetic recombination
• hand
• pathogenicity
• phylogeny
• whole-genome analysis

Hand, foot, and mouth disease (HFMD) is a common infectious disease occurring in children under 5 years of age worldwide, and Enterovirus A71 (EV-A71) and Coxsackievirus A16 (CVA-16) are identified as the predominant pathogens. In recent years, Coxsackievirus A6 (CVA-6) and Coxsackievirus A10 (CVA-10) have played more and more important role in a series of HFMD outbreaks. This study aimed to understand the epidemic characteristics associated with HFMD outbreak in Guangzhou, 2018.

The clinical and laboratory data of 1220 enterovirus-associated HFMD patients in 2018 were analysed in this study. Molecular diagnostic methods were performed to identify its serotypes. Phylogenetic analyses were depicted based on the complete VP1 gene.

There were 21 enterovirus serotypes detected in Guangzhou in 2018. Three serotypes of enterovirus, CVA-6 (364/1220, 29.8%), CVA-10 (305/1220, 25.0%), and CVA-16 (397/1220, 32.5%), were identified as the causative pathogens and accounted for 87.3% among all 1220 HFMD patients. In different seasons, CVA-6 was the predominant pathogen of HFMD during autumn, and CVA-10 as well as CVA-16 were more prevalent in summer. Patients infected by CVA-6, CVA-10 or CVA-16 showed similar clinical features and laboratory characteristics, and the ratios of severe HFMD were 5.8, 5.9, and 1.5% in the three serotypes. Phylogenetic analyses of VP1 sequences showed that the CVA-6, CVA-10, and CVA-16 sequences belonged to the sub-genogroup E2, genogroup E, and genogroup B1, respectively.

CVA-6, CVA-10, and CVA-16 were the predominant and co-circulated serotypes in Guangzhou China, 2018, which should be the new target for prevention and control of HFMD. Our findings provide useful information for diagnosis, treatment, and prevention of HFMD.

• Co-circulation
• Coxsackievirus
• Enterovirus
• Hand, foot, and mouth disease

• coxsackievirus A6
• hand-foot-mouth disease
• human enteroviruses
• onychomadesis

• biomarker
• foot and mouth disease
• hand
• scara3

• Biomarkers/metabolism
• Case-Control Studies
• Child, Preschool
• Female
• Hand, Foot and Mouth Disease/diagnosis
• Hand, Foot and Mouth Disease/genetics
• Hand, Foot and Mouth Disease/metabolism
• Heat-Shock Proteins/genetics
• Heat-Shock Proteins/metabolism
• Humans
• Infant
• Male
• RNA, Messenger/metabolism
• Scavenger Receptors, Class A/genetics
• Scavenger Receptors, Class A/metabolism
• Severity of Illness Index

• Adolescent
• Adult
• Age Distribution
• Aged
• Child
• Child, Preschool
• Cyprus/epidemiology
• Enterovirus/genetics
• Enterovirus Infections/epidemiology
• Female
• Genotype
• Humans
• Infant
• Infant, Newborn
• Male
• Middle Aged
• Molecular Epidemiology
• Phylogeny
• Young Adult

Hand, foot, and mouth disease (HFMD) is well recognized as one of the major threats to children’s health globally. The increasing complexity of the etiology of HFMD still challenges disease control in China. There is little surveillance of the molecular epidemiological characteristics of the enteroviruses (EVs) that cause HFMD in Neijiang city or the Sichuan Basin area in Southwest China. In this study, demographic and epidemiological information for 14,928 probable HFMD cases was extracted and analyzed to describe the epidemic features of HFMD in Neijiang city from Jan 2010 to Dec 2017. The swab samples of select probable HFMD cases from 2012 to 2017 were tested by reverse transcription (RT) real-time PCR to identify the serotype distribution of EVs, and 110 randomly selected RT-real-time PCR positive samples were then amplified and analyzed for the VP1 or VP4 regions of EVs to further analyze the phylogenetic characteristics of the circulating strains in this area. The eight-year average annual incidence was 49.82 per 100,000 in Neijiang. The incidence rates varied between 19.51 and 70.73 per 100,000, demonstrating peaks of incidence in even-number years (2012, 2014 and 2016). The median age of the probable cases was 27 months and the interquartile range (25^th to 75^th percentile) of ages for the probable HFMD cases was between 14 and 42 months. The male-to-female ratio of the probable HFMD cases was 1.47:1, and scattered children were the major population classification (81.7%). Two epidemic peaks were observed: one major peak between April and July and the other lesser peak between October and December. Of 6513 probable cases tested with RT-real-time PCR, 4015 (61.6%) were positive for enterovirus with the serotype distribution as follows: EV71+, 30.1% (n = 1210); CV-A16+, 28.7% (n = 1154) and a sole pan-enterovirus+, 41.1% (n = 1651). A total of 91 cases (82.7%, 91/110) were successfully amplified and underwent phylogenetic analysis: all EV71+ cases were C4a serotype (n = 23/30); all CV-A16+ cases were B2b serotype (n = 24/30); of 42 sole pan-enterovirus+ samples, 20 were CV-A6, 14 were CV-A10 and the rest within this group were CV-A4 (n = 4), CV-A8 (n = 2), CV-A9 (n = 1) and CV-B3 (n = 1). Our findings provide important evidence that aids the improvement of strategies for vaccination against HFMD and comprehensive disease control in China.

• coxsackievirus a6
• hand, foot, and mouth disease
• molecular epidemiology

• Base Sequence
• Capsid Proteins/genetics
• Child
• Child, Preschool
• China/epidemiology
• Disease Outbreaks
• Enterovirus/classification
• Enterovirus/genetics
• Enterovirus/isolation & purification
• Female
• Hand, Foot and Mouth Disease/epidemiology
• Hand, Foot and Mouth Disease/prevention & control
• Hand, Foot and Mouth Disease/virology
• Humans
• Infant
• Male
• Molecular Epidemiology
• Sequence Analysis, DNA

Hand, foot and mouth disease (HFMD) is a major public health issue in Asia and has global pandemic potential. Coxsackievirus A6 (CV-A6) was detected in 514/2,230 (23%) of HFMD patients admitted to 3 major hospitals in southern Vietnam during 2011–2015. Of these patients, 93 (18%) had severe HFMD. Phylogenetic analysis of 98 genome sequences revealed they belonged to cluster A and had been circulating in Vietnam for 2 years before emergence. CV-A6 movement among localities within Vietnam occurred frequently, whereas viral movement across international borders appeared rare. Skyline plots identified fluctuations in the relative genetic diversity of CV-A6 corresponding to large CV-A6–associated HFMD outbreaks worldwide. These data show that CV-A6 is an emerging pathogen and emphasize the necessity of active surveillance and understanding the mechanisms that shape the pathogen evolution and emergence, which is essential for development and implementation of intervention strategies.

• Asia
• Hand foot and mouth disease
• Vietnam
• coxsackievirus A6
• deep sequencing
• enteroviruses
• phylogeny
• viruses

• Adolescent
• Adult
• Child
• Communicable Diseases, Emerging/epidemiology
• Communicable Diseases, Emerging/virology
• Coxsackievirus Infections/epidemiology
• Coxsackievirus Infections/virology
• Enterovirus A, Human/classification
• Enterovirus A, Human/genetics
• Enterovirus A, Human/isolation & purification
• Female
• Genome, Viral
• Genomics/methods
• Hand, Foot and Mouth Disease/epidemiology
• Hand, Foot and Mouth Disease/virology
• Humans
• Male
• Phylogeny
• Phylogeography
• Vietnam/epidemiology
• Whole Genome Sequencing
• Young Adult

• Wellcome Trust
• 101104/Z/13/Z Wellcome Trust
• 106680/B/14/Z Wellcome Trust
• 204904/Z/16/Z Wellcome Trust

Coxsackievirus A2 (CV-A2) has emerged as an important etiological agent in the hand, foot, and mouth disease and herpangina pathogen spectrum because of its high global prevalence. In the present study, we investigated the evolutionary dynamics of CV-A2 circulating in China. We analyzed a total of 163 entire VP1 sequences of CV-A2, including 74 sequences generated from the present study and 89 sequences collected from the GenBank database. Phylogenetic analysis based on the entire VP1 nucleotide sequences confirmed the persistent circulation of the predominant genotype D in mainland of China since 2008. Cluster analysis grouped the sequences into two distinct clusters, clusters 1 and 2, with most grouped under cluster 2. After 2012, cluster 1 was gradually replaced by cluster 2. Results of Bayesian Markov chain Monte Carlo analysis suggested that multiple lineages of genotype D were transmitted in mainland of China at an estimated evolutionary rate of 6.32×10⁻³ substitutions per site per year, which is consistent with the global evolutionary rate of CV-A2 (5.82×10⁻³ substitutions per site per year). Continuous transmission and evolution of CV-A2 resulted in the genetic polymorphism.

• Capsid Proteins/genetics
• China
• Enterovirus/genetics
• Enterovirus/isolation & purification
• Evolution, Molecular
• Genotype
• Geography
• Humans
• Phylogeny

• National Key Technologies Research and Development Program of National Ministry of Science

• Coxsackievirus A6
• Epidemiology
• Hand-foot-and-mouth disease
• Hong Kong
• Novel
• Recombination

• Adolescent
• Child
• Child, Preschool
• China
• Enterovirus C, Human/genetics
• Evolution, Molecular
• Female
• Genotype
• Hand, Foot and Mouth Disease/epidemiology
• Hand, Foot and Mouth Disease/genetics
• Humans
• Male

• The Key Technologies Research and Development Program from the Ministry of Science and Technology (grant numbers: 2013ZX10004-202,2012ZX10004201-003).

Coxsackievirus A6 (CVA6) is one of the major agents to cause hand, foot and mouth disease (HFMD) outbreaks globally. The objective of this study is to investigate the epidemiologic and clinical manifestations of CVA6 outbreak, and thus guide the diagnosis and treatment of the disease, as well as disease prevention.

An HFMD outbreak in a kindergarten was reported to Shijingshan District Center for Disease Control and Prevention (SCDC) on November 2, 2015 in Beijing, China. Epidemiological investigation was conducted. We performed a nine-week follow-up study to collect and analyze the clinical manifestations of HFMD cases.

The outbreak yield 56 (15.7%) clinical diagnosed HFMD cases out of 357 registered children in the kindergarten with the mean age of 3.5 years old. This outbreak lasted for three days and ceased after initiating infectious disease controlling procedures, including periodical suspension of the kindergarten activities, environmental disinfection, and family health education. Fifty-one cases were followed for nine weeks. The positive rate of clinical manifestations of rash, fever, desquamation, pigmentation and onychomadesis were 100.0%, 84.3%, 68.6%, 17.6% and 43.1%, respectively. Children developed desquamation within the first 4 weeks after disease onset and developed onychomadesis between the 3th and 8th week after disease onset. Children with desquamation had 9.3 (95%CI: 1.836–47.437) times higher odds of developing onychomadesis compared to those without this manifestation. Ten out of 14 collected samples were CVA6 positive, and five positive samples shared a high degree of similarity in the VP1 nucleotide and amino acid sequences (99.9–100.0% and 100%).

This HFMD outbreak was caused by CVA6, featured with delayed symptoms. Emerging CVA6-associated HFMD and its delayed symptoms should be paid more attention to reduce outbreaks and provide more information to doctors and parents.

The online version of this article (10.1186/s12887-018-1253-1) contains supplementary material, which is available to authorized users.

• Coxsackievirus A6
• Disease outbreak
• Hand foot and mouth disease
• Onychomadesis

• Coxsackievirus A6
• neutralizing antibody
• pseudovirus
• pseudovirus luciferase assay

Hand, foot and mouth disease (HFMD) is usually caused by several serotypes from human enterovirus A species, including enterovirus 71 (EV-A71) and coxsackievirus A16 (CV-A16). Two inactivated monovalent EV-A71 vaccines have been recently licensed in China and monovalent CV-A16 vaccine and bivalent EV-A71 and CV-A16 vaccine are under development. Methods: Using notifications from the national surveillance system, we describe the epidemiology and dynamics of HFMD in the country, before the introduction of EV-A71 vaccination, from 2008 through 2015. Results: Laboratory-identified serotype categories, i.e. CV-A16, EV-A71 and other enteroviruses, circulated annually. EV-A71 remained the most virulent serotype and was the major serotype for fatal cases (range: 88.5–95.4%) and severe cases (range: 50.7–82.3%) across years. Except for 2013 and 2015, when other enteroviruses were more frequently found in mild HFMD (48.8% and 52.5%), EV-A71 was more frequently detected from mild cases in the rest of the years covered by the study (range: 39.4–52.6%). The incidence rates and severity risks of HFMD associated with all serotype categories were the highest for children aged 1 year and younger, and decreased with increasing age. Discussion/conclusion: This study provides baseline epidemiology for evaluation of vaccine impact and potential serotype replacement.

• HFMD
• case-fatality
• epidemiology
• periodicity

• and mouth disease
• coxsackievirus A6
• foot
• hand
• lethality
• mouse model
• pathogenicity
• recombination

• Coxsackievirus A6 (CV-A6)
• Epidemic
• Evolution analysis
• Hand, foot, and mouth disease (HFMD)
• Human enteroviruses (human EVs)

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

Aside from enterovirus 71 (EV71) and coxsackie virus A16 (CV-A16), viruses that are known to cause hand-foot-and-mouth disease (HFMD), epidemiological profiles of other enteroviruses that induce HFMD are limited. We collected 9949 laboratory surveillance HFMD cases and 1230 serum samples from infants and children in Shanghai from 2012–2016. Since 2013, CV-A6 has displaced EV71 and CV-A16 to become the predominant serotype. Interestingly, novel epidemiological patterns in EV71 and CV-A16 infections were observed, with one large peak in both 2012 and 2014, followed by two smaller peaks in the respective following years (2013 and 2015). Through sequencing, we found that C4a, B1b, D-Cluster-1 and B constituted the major subgenotypes of EV71, CV-A16, CV-A6 and CV-A10, respectively. Among healthy individuals, 50.49% and 54.23% had positive neutralising antibodies (NtAbs) against EV71 and CV-A16, respectively, indicating that EV71 and CV-A16 silent infections were common. These populations may be an important potential source of infection. The overall seropositive rate of EV71 NtAbs showed a fluctuating, markedly downward trend, indicating the potential risk of a future EV71 epidemic. High CV-A16 NtAb seroprevalence corroborated a documented CV-A16 ‘silent’ epidemic. Children aged 1–5 years had the lowest EV71 NtAb seropositive rate, whereas those aged 1–2 years exhibited the lowest CV-A16 NtAb seropositive rate. This is the first comprehensive investigation of the epidemiology and aetiology, as well as the seroprevalence, of HFMD in Shanghai between 2012 and 2016. This study provides the latest insights into developing a more efficient HMFD vaccination programme.

A total of 807 entire VP1 sequences of Coxsackievirus A6 (CV-A6) from mainland of China from 1992 to 2015, including 520 in this study and 287 from the GenBank database, were analysed to provide a basic framework of molecular epidemiological characteristics of CV-A6 in China. Sixty-five VP1 sequences including 46 representative CV-A6 isolates from 807 Chinese strains and 19 international strains from GenBank were used for describing the genotypes and sub-genotypes. The results revealed that CV-A6 strains can be categorised into 4 genotypes designated as A, B, C, and D according to previous data and can be further subdivided into B1–B2, C1–C2, and D1–D3 sub-genotypes. D3 is the predominant sub-genotype that circulated in recent years in mainland of China and represents 734 of 807 Chinese isolates. Sixty-six strains belong to D2, whereas B1 and C1 comprise a single strain each, and five AFP strains formed B2. Sub-genotype D3 first circulated in 2008 and has become the predominant sub-genotype since 2009 and then reached a peak in 2013, while D2 was mostly undetectable in the past years. These data revealed different transmission stages of CV-A6 in mainland of China and that sub-genotype D3 may have stronger transmission ability.

• coxsackievirus
• enterovirus
• hand-foot-mouth disease
• molecular epidemiology
• pathogen spectrum

• Capsid Proteins/genetics
• Child, Preschool
• China/epidemiology
• Disease Outbreaks
• Enterovirus/genetics
• Enterovirus/pathogenicity
• Female
• Genetic Testing
• Genotype
• Hand, Foot and Mouth Disease/epidemiology
• Hand, Foot and Mouth Disease/virology
• Humans
• Male
• Phylogeny
• Prevalence
• RNA, Viral/genetics
• Real-Time Polymerase Chain Reaction
• Serogroup

• Japan
• coxsackievirus A6
• hand, foot, and mouth disease

There are a variety of enteroviruses (EV) that can cause hand, foot and mouth disease (HFMD), and the major pathogens include enterovirus 71 (EV71) and coxasckievirus A16 (CVA16). EV71 and CVA16 have attracted much attention for their high prevalence and pathogenicity, and disease surveillance and vaccine development are mainly concentrated on them. EV71 can cause serious harm to children with HFMD, especially the damage to the nervous system such as aseptic meningitis, brain stem encephalitis and paralytic disease, or even lead to death. However, in recent years, due to the epidemic of EV71 and CVA16, people have established an immune barrier through natural infection in a certain degree. Although there is no cross protection between types, the immune protection against the relevant type can persist for a long time. Thus, the number of HFMD cases caused by EV71 and CVA16 shows a decreasing trend, while the epidemic of HFMD caused by other EV exhibits an upward trend. Further studies found that non-EV71 and non-CVA16 EV are very complex, and there are also differences in EV prevalence each year, which makes the development, evolution and control of HFMD become complicated. At present, there is no enough attention paid to the sporadic virus in the HFMD epidemic, and a complete research system for non-EV71 and non-CVA16 EV has not formed. Therefore, it is necessary to strengthen the monitoring of multiple non-EV71 and non-CVA16 EV, further investigate their pathogenicity and genetic characteristics, and evaluate the relative frequency and biological hazard of infection. In this review, we summarize a variety of EV changes, molecular evolution, as well as typical epidemics, which may provide clues to the development of antiviral drugs and vaccines, and prevention and control of HFMD.

• Hand, foot and mouth disease
• Epidemic situation
• Enterovirus
• Evolution trend

• VP1 gene
• enterovirus 71(EV-A71)
• hand, foot, and mouth disease (HFMD)
• phylogenetic analysis

• Bayesian method
• Coxsackievirus A16
• Genetic diversity
• HFMD
• Molecular evolution

• Coxsackievirus A6
• Pichia pastoris
• Vaccine
• Virus-like particle

• Coxsackievirus A6
• clinical features
• enterovirus 71
• epidemic
• hand foot and mouth disease