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Objective: To establish a new method for rapid detection of Coxsachie virus A16 (CA16) hand, foot and mouth disease pathogens based on fluorescence resonance energy transfer (FRET) technique, to evaluate the detection effect and to make the method to meet the requirements of large sample size detection during the outbreak of disease. Methods: Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis and bicinchoninic acid (BCA) protein assay were used to identify the purity of CA16 chicken yolk antibody (CA16-IgY) and the protein level. Indirect enzyme-linked immunosorbent assay (iELISA) was used to detect the titer and specificity of anti-CA16 IgY antibody. The size, morphology and characterization of gold nanoparticles (AuNPs) and their biological probes (IgY-AuNPs) were determined by UV-visible spectroscopy (UV-Vis), infrared spectroscopy (FTIR) and transmission electron microscopy (TEM). The CA16 detection system was constructed based on FRET technique. The sensitivity and specificity of the detection method and clinical sample detection were evaluated by optimizing the IgY-AuNPs concentration, sodium chloride (NaCl) dosage, fluorescence recovery time and other indicators. Results: The CA16-IgY had high purity, the titer was 1:128 000, the average protein level was 12. 15 mg · L-1, and CA16-IgY had good specificity. The results of UV-Vis, FTIR and TEM of AuNPs and IgY-AuNPs showed that IgY was successfully labeled onto the surface of AuNPs, which suggested that IgY-AuNPs could specially recognize CA16 was successfully prepared by electrostatic self-assembly. The CA16 detection system was constructed based on FRET technology, after optimization of the detection system, the optimal dosage of IgY-AuNPs was determined to be 0.52 X 10-3 g · L-1, the optimal dosage of NaCl was 40 μL and the optimal fluorescence recovery time was 90 min. The standard curve of the established detection method was I525 ntu= 15. 452 IgC-9. 746, R2 = 0.993 2, the detection limit was as 1 X 104 PFU · ml-1. Compared with qRT-PCR, the agreement rate reached 93. 75%. Conclusion: A new rapid detection method for CA16 hand, foot and mouth disease pathogens is successfully established, which can be applied to laboratory and clinical tests.
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Hand-foot-mouth disease (HFMD) is a common infectious disease caused by enterovirus in children. It has a high incidence and can cause fatal complications such as pulmonary edema, myocarditis and aseptic meningitis, seriously threatening the health of children. At present, some core problems such as the pathogenesis of disease, the relationship between different genotypes of pathogenic viruses, the pharmacodynamic evaluation methods, and the antiviral mechanism of drugs are still unclear. The construction of disease animal models with simulation performance of human exposure is the key to solve the above problems. Researchers both at home and abroad have established a variety of animal models for HFMD, of which enterovirus 71 (EV71) and coxsackievirus A16 (CA16) are most common and most widely used. Both EV71 and CA16 are enterovirus A in picornavirus family, so they have similarities in terms of pathogenicity, infection and replication characteristics, clinical symptoms caused by infection and immune response, but also have significant differences in age of susceptibility, method of infection, as well as neurotoxicity, clinical symptoms and signs, and degree of tissue and organ damage. Therefore, researchers shall select and establish proper animal models based on actual conditions, which is critical to the reliability of the results. In this paper, the different types of HFMD animal models established by EV71 and CA16 viruses were reviewed, especially on the species strains, virus strain types, infection methods, and characteristics of viral infections in each model, and the characteristics and clinical symptoms of HFMD induced by EV71 and CA16 were also investigated to provide reference for related research.
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Objective To analyze the epidemiological characteristics of hand foot and mouth disease (HFMD) cases caused by Coxsackie virus A16 (Cox A16) in Guangdong province from 2012 to 2016.Methods The data of mild HFMD cases caused by Cox A 16 were collected from 8 sentinel hospitals in 8 prefecture-level cities in Guangdong to estimate Cox A16 infection status and its population and time distribution characteristics.Results (1) The highest estimated incidence of Cox A16 infection was in 2014 (113.0/100 000),followed by 2016 (86.4/100 000) and 2012 (79.1/100 000),while the estimated incidence was lower in 2015 (29.0/100 000) and 2013 (28.8/100 000).(2) Cox A16 was confirmed to be the predominant pathogen causing HFMD outbreaks (54.6％,89/163).The number of outbreaks in the year with high incidence (28 outbreaks) was 11.2 times higher than that in the year with low incidence (2.5 outbreaks).(3) Across all age groups,the annual estimated incidence of Cox A16 infection decreased with age (trend x2=853 905.63,P＜0.01).The incidence was highest in age group 1 year (1 449.2/100 000),followed by that in age group 3 years (1 097.0/100 000),in age group 2 years (1 083.5/100 000),in age group 4 years (687.8/100 000) and in age group 0 year (604.9/100 000).Among the age groups ＜12 months,the estimated incidence increased with age (trend g2=5 541.77,P ＜ 0.01),which was highest in age group 11-months (2 105.1/100 000),followed by that in age groups 10-months (1 448.6/100 000),9-months (938.3/100 000),8-months (703.3/100 000) and 6-months (664.6/100 000).(4) The annual incidence peak was during May (143.9/100 000)-June (131.5/100 000).Conclusion The prevalence of Cox A16 infection differed with year in Guangdong during 2012-2016.When the incidence of Cox A16 infection was high,more outbreaks occurred.The prevalence occurred mainly in nurseries and kindergartens from May to June each year.Children aged 0-4 years were the high risk group for Cox A16 infection,children aged 6-11 months were at high risk for Cox A16 infection.
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Objective To analyze the epidemiological characteristics of hand foot and mouth disease (HFMD) cases caused by Coxsackie virus A16 (Cox A16) in Guangdong province from 2012 to 2016.Methods The data of mild HFMD cases caused by Cox A 16 were collected from 8 sentinel hospitals in 8 prefecture-level cities in Guangdong to estimate Cox A16 infection status and its population and time distribution characteristics.Results (1) The highest estimated incidence of Cox A16 infection was in 2014 (113.0/100 000),followed by 2016 (86.4/100 000) and 2012 (79.1/100 000),while the estimated incidence was lower in 2015 (29.0/100 000) and 2013 (28.8/100 000).(2) Cox A16 was confirmed to be the predominant pathogen causing HFMD outbreaks (54.6％,89/163).The number of outbreaks in the year with high incidence (28 outbreaks) was 11.2 times higher than that in the year with low incidence (2.5 outbreaks).(3) Across all age groups,the annual estimated incidence of Cox A16 infection decreased with age (trend x2=853 905.63,P＜0.01).The incidence was highest in age group 1 year (1 449.2/100 000),followed by that in age group 3 years (1 097.0/100 000),in age group 2 years (1 083.5/100 000),in age group 4 years (687.8/100 000) and in age group 0 year (604.9/100 000).Among the age groups ＜12 months,the estimated incidence increased with age (trend g2=5 541.77,P ＜ 0.01),which was highest in age group 11-months (2 105.1/100 000),followed by that in age groups 10-months (1 448.6/100 000),9-months (938.3/100 000),8-months (703.3/100 000) and 6-months (664.6/100 000).(4) The annual incidence peak was during May (143.9/100 000)-June (131.5/100 000).Conclusion The prevalence of Cox A16 infection differed with year in Guangdong during 2012-2016.When the incidence of Cox A16 infection was high,more outbreaks occurred.The prevalence occurred mainly in nurseries and kindergartens from May to June each year.Children aged 0-4 years were the high risk group for Cox A16 infection,children aged 6-11 months were at high risk for Cox A16 infection.
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Objective To investigate the etiological agents of the outbreak of hand,foot and mouth disease (HFMD) in Minhang District of Shanghai from 2009 to 2016,and to provide evidence for the prevention and management policy of HFMD by collecting suspected HFMD samples for laboratory testing from HFMD sentinel hospitals,the Children's Hospitals of Fudan University and jurisdiction community health service centers.Methods Specimens including stools,throat swabs and anal swabs were collected from patients suspected of HFMD from surveillance hospitals,i.e.,Children's Hospital of Fudan University and jurisdiction community health service centers during 2009 and 2016.The specimens were detected by real-time RT-PCR with the five types of viruses including panenterovirus (EV),enterovirus 71 (EV71),Coxsackie virus A16 (CVA16),Coxsackie virus A6 (CVA6) and Coxsackie virus A10 (CVA10).The distribution characteristics of pathogens were analyzed.Results During 2009 and 2016 we collected 3 744 cases of HFMD,the positive detection rate of pan-enterovirus were 84.83％ (3 176 cases),including EV71 (64.45％) and CVA16 (15.77％),then were CVA6 (9.23％) and other EV (8.78％),and CVA10 was only 0.76％.The major etiological agent was EV71.The advantage of different years and different season strain presented dynamic change,mainly EV71 and CVA16 co-popular in 2009,mainly EV71 epidemic from 2010 to 2011,EV71 and CVA16 were co-popular again in 2012,and the priority was EV71,then EV71 and other EV were co-popular in 2013.After 2014,CVA6 gradually increased,mainly CVA6 was popular 2015 and 2016,especially in 2016,the proportion of CVA6 ranged as high as 40.55％.CVA10 showed sporadic distribution trend in very low proportion.From the difference of age and gender in HFMD cases,the males were more than the females (1.68 ∶ 1),and the incidence was the highest in children of 1-3 years old.The peak of incidence occured in April to July,then in September to November.Conclusions HFMD onset seasons mainly were summer and fall-and-winter in bimodal popular models.It happens in children under 5 years of age.The incidence was higher in male than in female.The advantage of different years and different season strains present dynamic change,and the prevalence has obvious age and season limit.
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Objective To investigate the etiological agents of the outbreak of hand,foot and mouth disease (HFMD) in Minhang District of Shanghai from 2009 to 2016,and to provide evidence for the prevention and management policy of HFMD by collecting suspected HFMD samples for laboratory testing from HFMD sentinel hospitals,the Children's Hospitals of Fudan University and jurisdiction community health service centers.Methods Specimens including stools,throat swabs and anal swabs were collected from patients suspected of HFMD from surveillance hospitals,i.e.,Children's Hospital of Fudan University and jurisdiction community health service centers during 2009 and 2016.The specimens were detected by real-time RT-PCR with the five types of viruses including panenterovirus (EV),enterovirus 71 (EV71),Coxsackie virus A16 (CVA16),Coxsackie virus A6 (CVA6) and Coxsackie virus A10 (CVA10).The distribution characteristics of pathogens were analyzed.Results During 2009 and 2016 we collected 3 744 cases of HFMD,the positive detection rate of pan-enterovirus were 84.83％ (3 176 cases),including EV71 (64.45％) and CVA16 (15.77％),then were CVA6 (9.23％) and other EV (8.78％),and CVA10 was only 0.76％.The major etiological agent was EV71.The advantage of different years and different season strain presented dynamic change,mainly EV71 and CVA16 co-popular in 2009,mainly EV71 epidemic from 2010 to 2011,EV71 and CVA16 were co-popular again in 2012,and the priority was EV71,then EV71 and other EV were co-popular in 2013.After 2014,CVA6 gradually increased,mainly CVA6 was popular 2015 and 2016,especially in 2016,the proportion of CVA6 ranged as high as 40.55％.CVA10 showed sporadic distribution trend in very low proportion.From the difference of age and gender in HFMD cases,the males were more than the females (1.68 ∶ 1),and the incidence was the highest in children of 1-3 years old.The peak of incidence occured in April to July,then in September to November.Conclusions HFMD onset seasons mainly were summer and fall-and-winter in bimodal popular models.It happens in children under 5 years of age.The incidence was higher in male than in female.The advantage of different years and different season strains present dynamic change,and the prevalence has obvious age and season limit.
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Objective To explore the epidemiologic and clinical features of hand, foot and mouth disease (HFMD) caused by Coxsackie virus A16 (CA16) in Suzhou from 2010 to 2014, and analyze the relationship between the SNPs of oligoadenylate synthetase 1 (OAS1) and HFMD caused by CA16 infection.MethodsThe clinical data of children diagnosed with HFMD caused by CA16 during 2010 and 2014 were collected. The epidemiological characteristics were analyzed. Among them, 167 cases were selected to make comparison of the clinical features with 166 cases of HFMD caused by EV71 infection in the same period. The genotyping ofOAS1 rs10774671 was detected by TaqMan probe technique in 167 cases of CA16 infection children, 166 cases of EV71 infection children with HFMD and 163 healthy children. The relationship between polymorphism of gene and infection of CA 16 was analyzed.ResultsA total of 9 016 children with HFMD were included. CA16 nucleic acid detected to be positive in 762 cases. The detection rate was 8.45%. CA16 infection was most commonly in summer. Children under 5 years old accounted for 94.62% infected. Compared with EV71 infected children, CA16 infected children had shorter fever time, severer oral herpes, ulcer, and rash in hand, foot and hip, lesser nervous system involvement, fewer cases of high lactate dehydrogenase, high C reactive protein, high IgM or IgG, and signiifcant changes in the percentage of CD3+, CD3+CD4+, CD3+CD8+ and CD3-CD19+ (P all?
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Objective To understand the infection status of enterovirus 71(EV71) and coxsackievirus A16(Cox A16) among children receiving health examination for child care setting entrance in Beijing and their related medical care seeking practice and provide evidence for the estimation of disease burden caused by hand foot and mouth disease(HFMD). Methods Serological survey was conducted in the local children receiving health examination for child care setting entrance. Enzyme-linked immunosorbent assay (ELISA) was conducted to detect anti-EV71 and anti-Cox A16 IgG and IgM. Results A total of 813 children were surveyed(mean age:3.5±1.0 year old). The seropositive rate was 61.9%and 4.4%for anti-Cox A16 IgG and IgM. The seropositive rate was 9.3% and 1.1% for anti-EV71 IgG and IgM. No significant difference was observed in sex specific seropositive rate (P>0.05). However,significant differences were found in seropositive rate among different age groups(P<0.05). Among the children who were anti-Cox A16 positive, 7.8%had ever had rashes on their hands and feet,mouth or buttocks(HFMD-like rashes). Among the children who were anti-EV71 positive,10.7%had ever had HFMD-like rashes. For the children who were anti-Cox A16 or anti-EV71 positive,only 7.1% were brought to see doctors by their parents. However,among the seropositive children with rashes,80.5% were brought to see doctors. Conclusion In the healthy children at the age to go to child care setting in Beijing,most had ever infected with Cox A16. The anti-EV71 positive rate was much lower than the anti-Cox A16 positive rate. It was necessary to strengthen the prevention and control of EV71 infection in child cares settings.
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Objective To understand the infection status of enterovirus 71(EV71) and coxsackievirus A16(Cox A16) among children receiving health examination for child care setting entrance in Beijing and their related medical care seeking practice and provide evidence for the estimation of disease burden caused by hand foot and mouth disease(HFMD). Methods Serological survey was conducted in the local children receiving health examination for child care setting entrance. Enzyme-linked immunosorbent assay (ELISA) was conducted to detect anti-EV71 and anti-Cox A16 IgG and IgM. Results A total of 813 children were surveyed(mean age:3.5±1.0 year old). The seropositive rate was 61.9%and 4.4%for anti-Cox A16 IgG and IgM. The seropositive rate was 9.3% and 1.1% for anti-EV71 IgG and IgM. No significant difference was observed in sex specific seropositive rate (P>0.05). However,significant differences were found in seropositive rate among different age groups(P<0.05). Among the children who were anti-Cox A16 positive, 7.8%had ever had rashes on their hands and feet,mouth or buttocks(HFMD-like rashes). Among the children who were anti-EV71 positive,10.7%had ever had HFMD-like rashes. For the children who were anti-Cox A16 or anti-EV71 positive,only 7.1% were brought to see doctors by their parents. However,among the seropositive children with rashes,80.5% were brought to see doctors. Conclusion In the healthy children at the age to go to child care setting in Beijing,most had ever infected with Cox A16. The anti-EV71 positive rate was much lower than the anti-Cox A16 positive rate. It was necessary to strengthen the prevention and control of EV71 infection in child cares settings.
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OBJECTIVES: This study aimed to review etiological and epidemiological data for hand, foot, and mouth disease (HFMD) cases that occurred between 2008 and 2010 in Guangzhou City, to help develop and implement precautionary measures applicable for future outbreaks. METHODS: The characteristics of 4,753 HFMD episodes were retrospectively reviewed in 4,636 patients reported between 2008 and 2010 by the Guangdong Women and Children's Hospital, which is the national enterovirus monitoring agent and the designated hospital in China for treating severe HFMD. RESULTS: Out of 4,753 incident episodes reviewed, 525 patients were hospitalized; 60% were males. Most patients (93.8%) were children under 5 years old, with a median age at onset of 2.4 years. HFMD incidence peaked in April/May and September/October. From the total, 1,067 (22.4%) infections were positive for human enterovirus 71 (HEV71), 1,094 (23.0%) were positive for coxsackievirus A16 (CA16), and 941 (19.8%) were positive for other common enteroviruses. In contrast, 1,666 (35.0%) cases were negative to HEV71, CA16, and other common enteroviruses. Cross-correlation coefficients demonstrated associations between the number of cases, seasonal temperatures, and humidity. Among hospitalized cases, HEV71 was positive in 261 (24.5%), and 42 (3.9%) critical cases were positive for HEV71. CONCLUSION: Seasonal fluctuations and HEV71 and CA16 were the two key factors influencing the Guangzhou HFMD epidemic. The infection predominantly affected children younger than 5 years old.
Sujets)
Enfant d'âge préscolaire , Femelle , Humains , Nourrisson , Mâle , Syndrome mains-pieds-bouche/épidémiologie , Syndrome mains-pieds-bouche/virologie , Chine/épidémiologie , Incidence , Études rétrospectives , Saisons , Indice de gravité de la maladieRésumé
Objective To investigate the application and correlation of enterovirus 71(EV71)and Coxsackie virus A16(CA16)antibody screening in the hand,foot and mouth disease(HFMD).Methods The method of enzyme linked immunosorbent assay was used to test the serum antibody of EV71-IgM and CA16-IgM in 281 patients(142 male and 139 female)with HFMD.Results The positive rate of EV71-IgM antibody was 51.2％(144/281),the positive rate of CA16-IgM antibody was 32.0％(90/281).Seventeen patients(6.0％,17/281)had dual positive of EV71-IgM and CA 16-IgM antibody.The total detection rate was 77.2％(217/281).The positive rate of EV71-IgM and CA16-IgM antibody had significant difference(x2 =21.35,P ＜ 0.01).The positive rate of EV71-IgM and CA16-IgM antibody in male had significant difference compared with female[95.1％(135/142)vs.71.2％(99/139),P＜ 0.01].Conclusions At the beginning of the illness should use the joint detection of EV71-IgM and CA16-IgM antibody diagnose HFMD.It has significance in diagnosing and monitoring.
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Objective To understand the pathogen-carrying status of hand-foot-mouth disease (HFMD) among healthy people in Guangdong province.Methods Stool specimens were collected randomly on 7 age groups from 7 cities in Guangdong province.Real-time RT-PCR was used to detect enterovirus (EV),enterovirus 71 (EV71) and coxsackie virus A16 (CA16).Results Altogether,1285 stool specimens were collected.The positive rates of EV71,CA16 and other enterovirus were 0.39％ (5/1285),0.23％ (3/1285) and 7.00％ (90/1285),respectively.The highest EV71 positive rate (1.79％) was among the 4-6-year-old group,followed by the age group 0-3 with positive rate as 0.67％.EV71 was not found among the rest age groups.The highest CA16 positive rate (1.35％) was among the 4-6 year-olds group,but the CA16 was not found among the rest age groups.EV71 was only found among native population,with the positive-rate as 0.47％.CA16-positive rate was 0.19％ among the native population and 0.85％ among floating population,with no significant difference found (P＞0.05).The EV71 positive rate was 0.36％ among rural residents and 0.54％ among urban residents,but with no significant difference (P＞0.05).All CA16 was found among the urban residents.Conclusion Recessive infection of EV71 and CA16 were only found among 0-6 year-old group but not found among other groups,which suggested that the approaches on prevention and control should be targeted to all children especially on pre-school children.
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Objective To identify the etiology of 8 human hand-foot-mouth disease(HFMD)outbreaks in Beijing,during May to July 2007.Methods Reverse transcription-polymerase chain reaction(RT-PCR) method was used to directly type the specimens including fluid from the herpes and throat swabs from the HFMD patients.Using RD cell lines,the collected stool specimens were cultured followed by typing.Partial VP1 region of selected EV positive specimens and cultures were sequenced and both nucleic acid sequence and predicted amino acid sequence were analyzed.Results The two HFMD outhreaks in Daxing region in Beijing in 2007 were caused by enterovirus 71 type(EV71),and the others were caused by Coxsackie virus A16(Cox A16).Two EV71 strains caused epidemics in Daxing region in 2007 belonged to C4 subgenotype but on different branches in VP1 gene phylogenefic tree.The differences on nucleic acid sequence and amino acid sequence were 3.7% and 0.8% between the two EV71 stains.respectively.The Cox A16 strain in Shunyi region and the other strains were on different branches in phylogenetic tree,and the difference on nucleic acid and amino acid sequence were 3.7% and 0% respectively between the two Cox A16 shams.Conclusion The HFMD outbreaks occurred in Beijing in 2007 were caused mainly by EV71 and Cox A16, and there were two individual epidemic virus strains.Cox A16 seemed to spread more widely than EV71 in Beiiing,2007.
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Objective To develop a rapid, accurate, specific method to detect causative agent of hand, foot and mouth disease (HFMD). Methods Specific primers and probe were designed based on highly conserved VP1 region of enterovirus 71, coxsackie virus A16 and enterovirus. The sensitivity and specificity of the real-time RT-PCR was evaluated with 35 stool samples collected from pediatric patients with suspected HFMD and 20 clinical samples from health pediatric patients. Results Out of 35 clinical samples from suspected HFMD, 35 samples were identified as positive for enterovirus, 25 clinical samples were identified as positive for enterovirus 71, 8 clinical samples were identified as positive for coxsackie virus A16, among which 3 clinical samples were identified as positive for enterovirus 71 and coxsackie virus A16. The clinical diagnostic accordance rate is 85.71%. Out of 20 clinical samples from normal pediatric patients, 5 clinical samples were identified as positive for enterovirus, 20 clinical samples were negative for enterovirns 71 and coxsackie virus AI6. Conclusion Our results indicate real-time RT-PCR offers a rapid, sensitive, specific and cheap method to detect pathogen of HFMD from clinical specimens.