Evolutionary analysis of the VP1 gene of Echovirus 30 in Yunnan Province, China / 国际生物医学工程杂志

Objective:To study the genetic characteristics and genetic evolution of echovirus 30 (ECHO30) isolates in Yunnan Province, China.Methods:Virus isolation was performed on nucleic acid-positive samples for hand, foot, and mouth disease pathogen surveillance in Yunnan Province, and VP1 gene sequencing was performed. The sequences of eight ECHO30 isolates from Yunnan Province and the gene sequences of the VP1 region of the ECHO30 reference strain downloaded from GenBank were compared and analyzed using MEGA 5.0 software, and then a phylogenetic tree was constructed to measure the homology of nucleotides and amino acids between the isolates.Results:The ECHO30 virus was distributed in Wenshan, Qujing, Chuxiong, and Kunming in Yunnan Province. The ECHO30 virus was relatively common in Wenshan. ECHO30 isolates belonged to the H2 subtype of the H genotype, which was close to the local reference strain LC120939 in Yunnan Province. On the VP1 gene at site 5, the amino acid change ratio was more active, the amino acids were diverse, and mutations also occurred at sites 54, 156, 258, and so on. Nucleotide and amino acid homology were 84.0% - 100.0% and 98.4% - 100.0%, respectively.Conclusions:ECHO30 isolates from Yunnan Province have certain geographical characteristics and belong to H2 of the H genotype. The nucleotide differences in virus sequences among subtypes are small and have a close genetic relationship.

Human enteroviral infection in fibromyalgia: a case-control blinded study

SUMMARY OBJECTIVE: This study aimed to test the hypothesis that fibromyalgia is associated with a human enteroviral infection. METHODS: Venous peripheral blood samples from 27 patients fulfilling the American College of Rheumatology revised diagnostic criteria for fibromyalgia and from 26 age- and sex-matched controls, who underwent immunofluorescence assays for coxsackievirus A7 IgG, coxsackievirus B1 IgG, coxsackievirus A7 IgA, coxsackievirus B1 IgA, echovirus IgG, and echovirus IgA. These immunological tests were performed blind to group status. RESULTS: There were no significant differences between the patient and control groups in respect of positive results for coxsackievirus A7 IgG (p=0.467), coxsackievirus B1 IgG (p=0.491), coxsackievirus A7 IgA (p=0.586), coxsackievirus B1 IgA (p=0.467), echovirus IgG (p=0.236), and echovirus IgA (p=1). CONCLUSIONS: The results of this systematic study do not support the hypothesis that fibromyalgia is associated with infection by a human enterovirus.

Analysis of pathogen detection results of neonatal Echovirus 11 infection outbreaks in Guangdong Province in 2019 / 中华微生物学和免疫学杂志

Objective:To analyze the laboratory test results of two outbreaks of neonatal enterovirus infections in Guangdong Province in 2019 and the genetic characteristics of Echo11, aiming to provide reference for the prevention and control of neonatal enterovirus infections.Methods:The pathogenic specimens of neonatal cases suspected of enterovirus infection were collected. Fluorescence quantitative PCR and sequencing were used for enterovirus typing and identification, and virus isolation was carried out for positive specimens.The complete sequences of VP1 of Echo11 were amplified and sequenced and the phylogenetic analysis was performed using the bioinformatics software such as Danstar6, Bioedit7.09 and MEGA6.06.Results:A total of 93 specimens from 36 neonatal cases were collected. After identification, 55 specimens from 24 cases were positive for enterovirus, of which 23 cases were positive for Echo11 and one case was positive for Coxsackievirus B4(CVB4). A total of 29 enterovirus strains were isolated from the specimens of 19 cases, of which 28 were Echo11 from 18 cases, and one was CVB4. Phylogenetic analysis revealed that the nucleotide homology between the 18 strains of Echo11 in this study was 98.2%-100.0%, and the nucleotide homology between the Echo11 strains causing the two neonatal infections was 99.7%-100.0% and 99.8%-100.0%, respectively. Echo11 could be divided into six genotypes as A, B, C, D, E and F, in which genotype A and genotype C were further divided into A1-5 and C1-4, and genotype D could be divided into D1-5. The 18 strains of Echo11 in this study were all subtype D5.Conclusions:In 2019, two outbreaks of neonatal infections in medical institutions in Guangdong Province were caused by Echo11, which belonged to the genotype D5.

The rare enterovirus c99 and echovirus 29 strains in Brazil: potential risks associated to silent circulation

Human enteroviruses (EVs) are associated with a wide spectrum of human diseases. Here we report the complete genome sequences of one EV-C99 strain and one E29 strain obtained from children suffering from acute gastroenteritis, without symptoms of enteroviral syndromes. This is the first report of EV-C99 in South America, and the second E29 genome described worldwide. Continuous surveillance on EVs is vital to provide further understanding of the circulation of new or rare EV serotypes in the country. The present study also highlights the capacity of EVs to remain in silent circulation in populations.

Genetic characteristic analysis of the VP1 gene of echovirus 30 isolated from viral meningitis cases in Yunnan province, 2010-2013 / 中华实验和临床病毒学杂志

Objective@#In this study we analyzed the genetic characteristics of echovirus 30 (E-30) VP1 gene sequences from Yunnan province isolated from viral meningitis (VM) cases in 2010-2013.@*Methods@#RT-PCR and VP1 gene sequencing were done for 9 E-30 strains isolated from VM cases in 2010-2013. VP1 gene sequences of E-30 reference strains were downloaded from the GenBank and their nucleotide (nt) and amino acid (aa) diversities were calculated by MEGA 5.1 software, the phylogenetic tree was constructed and the genetic characteristics and molecular epidemiology were analyzed.@*Results@#In 2010-2013, 9 strains of E-30 viruses were detected from 79 VM cases caused by echoviruses, accounting for 11.39%(9/79), the overall positive rate was 1.63%(9/553). Phylogenetic analysis revealed that E-30 strains can be divided into four genotypes (genotype A, B, C and D), and genotype D can be further divided into seven sub-genotypes. Nine Yunnan VM isolates were distributed in D7 sub-genotype, and can be further clustered into 3 branches: 5 strains isolated in 2010 were clustered in branch 1, it is evident that these viruses were responsible for an aseptic meningitis outbreak in Kunming in that year; one 2011 isolate, together with 2013 isolate and one isolate from healthy children in 2010 were clustered in branch 2, these two branches were Yunnan special branches, and two 2011 isolates had the highest homology with 2003 VM outbreaks′ strains isolated from Shandong, Jiangsu, and Zhejiang, showing that these strains may have the same evolutionary sources.@*Conclusions@#Nine Yunnan VM isolates were distributed in D7 sub-genotype, and these strains have different evolutionary sources, showing that at different times E-30 viruses in the same sub-genotypes branch might prevail in different areas.

Antiviral Activity of Gemcitabine Against Echovirus 30 Infection in Vitro

Echovirus 30 is one of the major causes of meningitis in children and adults. The purpose of our current study was to investigate whether selected antiviral drugs could provide antiviral activity against echovirus 30. Using RD cells, we assessed the cytopathic effect of echovirus 30, including viral RNA levels as indicators of viral replication. The effects of gemcitabine were compared to rupintrivir, a well-known antiviral drug. To understand the activity gemcitabine exerts on the viral life cycle, time course and time-of-addition assays were implemented. The most effective compounds against echovirus 30 were gemcitabine and rupintrivir, as demonstrated by their concentration-dependent activity. Gemcitabine affects the early stages of echovirus 30 infection by disrupting viral replication. However, gemcitabine failed to directly inactivate echovirus 30 particles or impede viral uptake into the RD cells. Gemcitabine can be considered as a lead candidate in the development of echovirus 30 antiviral drugs, specifically in the early stages of echovirus 30 replication.

Molecular detection and sequence analysis of enteroviruses isolated from a suspected outbreak of aseptic meningitis / 中华微生物学和免疫学杂志

Objective To detect and analyze enteroviruses causing suspected aseptic meningitis in a kindergarten in Jinhua City, Zhejiang Province. Methods Viral RNA was extracted from samples and cDNA was prepared by reverse transcription. PCR was performed to amplify the partial sequences of the 5′-untranslated region ( UTR) and VP1 gene of enteroviruses. Serotypes of the viruses were determined by com-paring the homology between the partial sequences of VP1 gene. Phylogenetic tree of the partial VP1 se-quences was constructed using MEGA6. Results This study included seven patients and twenty-six asymp-tomatic students. Coxsakievirus A10 (CV-A10) was detected in 48. 5％ of the students and echovirus 6 (Echo 6) in 21. 2％. Besides, 12. 1％ of the students might be co-infected by the two viruses. Among the seven patients, six were infected by CV-A10 and the other one might have co-infection. According to the phylogenetic analysis, CV-A10 strains detected in this study were closely related to those isolated in China in recent years, including the strains isolated in Xiamen in 2015 and Yunnan in 2017, while the Echo 6 strains were phylogenetically related to those isolated in Yunnan, Guangzhou and Shandong in 2014. Conclusions CV-A10 and Echo 6 were detected in the cases with suspected aseptic meningitis and had close phylogenetic relationships to the strains appeared in China in recent years.

Genotype distribution of human enteroviruses isolated from swage in Shanghai during year 2013-2014 / 中华预防医学杂志

Objective@#To explore the time and genotype distribution of human enterovirus (HEV) isolated from sewage in Shanghai in 2013-2014.@*Methods@#One sewage sample each was collected from two local sewage plants located in Minhang District and Jiading District on the same day at the day 24-28 of every month from 2013 to 2014. Each sample weighed 1 L. The specimens were concentrated by anionic membrane absorption, eluted with beef extract solution, and then used to inoculate RD, HEp-2, and L20B cell lines. A total of 249 enterovirus strains were isolated from sewage samples during the study period, including 185 non-polio enterovirus (NPEV) and 64 poliovirus (PV) strains, which were identified as vaccine strains. RT-PCR and Sanger sequencing were performed to identify HEV genotypes. Homologous analysis of VP1 sequences was conducted using BioEdit (version 7.0.0). Phylogenetic analysis was performed using the neighbor-joining method based on the alignment of VP1 gene sequences using MEGA (version 4.0.2).@*Results@#Among 185 NPEV strains, 178 strains were successfully sequenced and classified into 15 genotypes, including coxsackievirus group B (CVB) 2, 3, and 5; enteric cytopathic human orphan (ECHO) virus 1, 3, 6, 7, 11, 13, 19, 20, 24, 25, and 30; and coxsackievirus group A 4. CVB5 and ECHO6 genotypes accounted for 33.5% (56 strains) and 24.9% (43 strains) of NPEV isolates, respectively. During the study period, HEV isolates were mainly isolated in summer and autumn in Minhang District. ECHO6 strains were frequently isolated from June 2013 to July 2014. Thereafter, the number of ECHO6 strains gradually reduced in the second half of 2014. CVB5 strains demonstrated scattered distribution from 2013 to the first half of 2014 and gradually increased in the second half of 2014. The distribution of ECHO6 and CVB5 strains in Jiading District was similar to that in Minhang District. In 2013-2014, CVB5 strains comprised C6 and C8 subgenotypes, which belong to two transmission chains and show large differences compared with foreign strains isolated during the same period. ECHO6 strains comprised C6, C8, and D9 subgenotypes, which belong to three transmission chains. Moreover, ECHO6 subgenotype D9 was a dominant subgenotype in Shanghai, with broad geographical distribution both at home and abroad.@*Conclusion@#Poliovirus was identified as a vaccine strain in environmental surveillance from June 2013 to April 2014 in Shanghai. Several transmission strains of ECHO6 and CVB5 were identified, which were the dominant serotypes.

Neurologic Manifestations according to Serotypes of Enterovirus in Pediatric Inpatient in Incheon

PURPOSE: Enterovirus infection in children can manifest various disease and enterovirus have many serotypes. This study was aimed to investigate neurologic manifestations according to serotypes of enterovirus in pediatric inpatients in Incheon. METHODS: We collected the stool samples from the admitted pediatric patients in Inha University Hospital from January 2015 to September 2016. Enterovirus detection and serotypes identification were performed by real-time reverse transcriptase polymerase chain reaction (RT-PCR) and semi-nested RT-PCR. RESULTS: A total of 527 samples were collected during study period and 170 patients (32.2%) were diagnosed with enterovirus infections. Genetic sequences of enteroviruses were identified: echovirus 18 (50, 40.5%), enterovirus 71 (12, 9.6%), coxakievirus A10 (10, 8.0%), echovirus 6 (7, 5.6%). Virus in patient with meningitis were identified: echovirus 18 (15, 75%), coxakievirus B5 (2, 10%), enterovirus 71 (2, 10%), and echovirus 6 (1, 5%). Neurologic manifestations of echovirus 18 are headache (15, 30%), vomiting (17, 34%), meningeal irritation sign (10, 20.0%). And enterovirus 71 have headache (3, 25%), vomiting (3, 25%), meningeal irritation sign (2, 16.0%), seizure (1, 8.3%), neurologic sequelae (1, 8.3%). Echovirus 18 and neurologic manifestation have a statistically significant correlation with other serotypes (r=0.701, P < 0.01) CONCLUSION: Echovirus 18 infection was more prominent in neurological symptoms than in other serotypes. The major serotype of meningitis was echovirus 18 but there was no reported neurologic sequelae. Enterovirus infection has different neurological symptoms, depending on the serotypes.

Echovirus serotypes circulating in Malaysia from 2002 to 2013

Objective: To identify the circulating serotypes of human echovirus in Malaysia from 2002 to 2013. Methods: A total of 31 retrospective samples from non-polio acute flacid paralysis, hand-food-and-mouth disease, viral meningitis and enterovirus cases were subjected to amplification of partial VP1 gene by RT-PCR. Results: Sequencing and phylogenetic analysis of the partial sequences identified presence of human echovirus and human coxsackie viruses. It was found that echovirus 11 was the commonly circulating serotype followed by echovirus 6, echovirus 7, echovirus 3, echovirus 9, echovirus 30 and echovirus 1 in decreasing order. Additionally two types of human coxsackie virus isolates were detected which were coxsackie A24 and B3. Conclusions: From the findings, there is a possibility that echovirus 11 is the predominant serotype among Malaysian patients with echovirus infection. However, a larger sample size will yield a more confident result to support this evidence.

Surveillance and genetic characteristics of echovirus 6 strains isolated from environmental sewage and aseptic encephalitis cases / 中华微生物学和免疫学杂志

Objective To analyze the genetic characteristics of echovirus 6 ( E-6) strains isolated from patients with acute meningitis/encephalitis syndrome ( AMES) in 2014 and sewage samples in 2013—2014 in Shandong province and to investigate their correlations.Methods Enterovirus strains were isolated from cerebrospinal fluid, stool and throat swab samples collected from 940 cases of AMES and 96 sewage samples used for environmental surveillance.The positive isolates were identified by molecular typing meth-od.Homologous and phylogenetic analyses based on the VP1 sequences of E-6 isolates were performed.Re-sults Altogether 47 E-6 strains were isolated from patients with AMES in 2014, accounting for 29.56%of all isolated enteroviruses ( EVs) strains.No E-6 strains were isolated from AMES cases in 2013.Data of the environmental surveillance showed that E-6 virus strains had been frequently detected in sewage samples since the summer of 2013 to the end of 2014.In total, 40 E-6 virus strains were isolated (7.87% of total isolated EVs strains) in 2013 and 139 E-6 virus strains (26.18%) in 2014.Phylogenetic analysis indicated that the E-6 isolates recruited in this study belonged to clusters A and C with high intracluster sequence iden-tities between AMES and environmental isolates.The nucleotide identities were 98.3%-100% among cluster A E-6 virus strains isolated from AMES cases in 2014 and 96.6%-100% among cluster A E-6 virus environ-mental isolates during the surveillance year 2013—2014.The cluster A E-6 virus strains shared 97.1%-100% nucleotide identities between the AMES and environmental isolates.For cluster C E-6 virus strains, the nucleotide identities were 100%, 98.7%-100% and 99.1%-100%, respectively.Conclusion The epidemic of viral encephalitis in Shandong province in 2014 was associated the transmission of two lineages of E-6 virus.Environmental surveillance revealed the potential epidemic of E-6 virus even before the epidemic of viral encephalitis in Shandong province, indicating the possibility of using environmental surveillance for early warning of related diseases.

Echovirus serotypes circulating in Malaysia from 2002 to 2013

OBJECTIVE@#To identify the circulating serotypes of human echovirus in Malaysia from 2002 to 2013.@*METHODS@#A total of 31 retrospective samples from non-polio acute flacid paralysis, hand-food-and-mouth disease, viral meningitis and enterovirus cases were subjected to amplification of partial VP1 gene by RT-PCR.@*RESULTS@#Sequencing and phylogenetic analysis of the partial sequences identified presence of human echovirus and human coxsackie viruses. It was found that echovirus 11 was the commonly circulating serotype followed by echovirus 6, echovirus 7, echovirus 3, echovirus 9, echovirus 30 and echovirus 1 in decreasing order. Additionally two types of human coxsackie virus isolates were detected which were coxsackie A24 and B3.@*CONCLUSIONS@#From the findings, there is a possibility that echovirus 11 is the predominant serotype among Malaysian patients with echovirus infection. However, a larger sample size will yield a more confident result to support this evidence.

Meningoencefalitis por echovirus30 complicada: reporte de dos casos / Complicated meningoencephalitis caused by echovirus30: report of two cases

Introducción: entre los principales virus causales de meningoencefalitis de evolución benigna en el 95 % de los niños están los enterovirus. Ante la infrecuencia de la gravedad de estos procesos, presentamos dos infantes complicados a Echovirus 30 recluidos en la unidad de terapia intensiva del Hospital Pediátrico Provincial Docente Eduardo Agramonte Piña de Camagüey, con el objetivo de mostrar la capacidad de estos agentes de producir daños severos del sistema nervioso central. Casos clínicos: se presentaron niños de 11 meses y un año de edad respectivamente ambos remitidos del municipio de Florida en la provincia de Camagüey que presentaron fiebre, vómitos, somnolencia, astenia y anorexia; por lo que se les realizó punciones lumbares con resultados de aumento de las células a predominio linfocitario, además uno de ellos presentó graves trastornos de perfusión, edema cerebral severo, y leucograma con granulaciones tóxicas; el otro paciente a las 72 horas de su ingreso comenzó con irritabilidad, ataxia y movimientos incordinados de los brazos; por lo que se le diagnosticó una cerebelitis con pronóstico reservado. El estudio virológico de las heces de ambos enfermos realizado en el Instituto de Medicina Tropical en La Habana, arrojó ECHO virus 30. Los dos enfermos tuvieron estadía hospitalaria de 15 y 20 días respectivamente con diagnóstico de meningoencefalitis viral. Conclusiones: los casos expuestos nos muestran el potencial de los enterovirus como el ECHO 30 de producir meningoencefalitis de evolución seria e impredecible, aunque nuestros pacientes resolvieron sin secuelas, es importante la vigilancia mantenida hacia los enterovirus en la población infantil para prevenir las infecciones del sistema nervioso central.

Introduction: enteroviruses are among the main causal viruses of meningoencephalitis of mild evolution in the 95 % of children. Given the infrequency of the seriousness of these complaints, the cases of two infants complicated with echovirus 30 are presented. The patients were confined to the intensive care unit of the Eduardo Agramonte Piña Teaching Provincial Pediatric Hospital. The cases are presented with the objective of showing the capacity of these agents of producing severe damage in the central nervous system. Clinical cases: the cases of two infants of 11 months and one year old are presented; both infants were brought about a remission of the Florida municipality, province of Camagüey. Both patients presented vomits, sleepiness, asthenia, and anorexia; because of that, they underwent a lumbar puncture the results of which were as follow: increase of the cells with lymphocytic predominance. One of the patients also presented serious disorders of perfusion, severe brain edema and the leucograma produced the presence of toxic granulations; the other patient started presenting irritability, ataxia, and uncoordinated movements of the arms 72 hours after being admitted and was diagnosed with cerebellitis with a reserved prognosis. The virology study of the feces of both patients was made in the Tropical Medicine Institute in Havana; the study produced the following result: echovirus 30. The patients had a hospital stay of 15 and 20 days respectively and were diagnosed with viral meningoencephalitis. Conclusions: the cases presented show us the potential of enteroviruses like ECHO 30 of producing meningoencephalitis of serious and unpredictable evolution. Although the patients recovered without aftereffects, a maintained vigilance on enteroviruses in children is important to prevent infections in the central nervous system.

Complete nucleotide sequence of a human echovirus 30 strain KM/A363/09 isolated in Yunnan, China in 2009 / 中华微生物学和免疫学杂志

Objective To analyze the genetic characteristics of the complete genome of a human echovirus 30 (Echo30) KM/A363/09 strain isolated in Yunnan, China in 2009.Methods Primers specif-ic for Echo30 were designed .The extracted RNA was amplified by using RT-PCR.Seven fragments covering the complete viral genome were sequenced and the complete sequences were aligned with other sequences of enterovirus reference strains downloaded from Genbank . By using Mega5.1, Geneious, RDP3 and SimPlot3.5.1 softwares, the phylogenetic and recombination analysis were carried out .Results The com-plete nucleotide sequence of KM/A363/09 isolate was 7425 bp in length, encoding 2194 amino acids.KM/A363/09 isolate was highly similar with Bastianni prototype strain showing the homology of 81.2%in nucle-otide and 95.8%in amino acid.The eight Echo30 isolates shared 81.2%-88.6%homologies in nucleotide sequences and 95.8%-97.8%in amino acid sequences .Phylogenetic analysis showed that the KM/A363/09 strain belonged to one clade of Echo 30 in China.The genetic recombination of KM/A363/09 isolate was detected in the non-structural region .Conclusion KM/A363/09 isolate belongs to one clade of Echo 30 in China indicating that the evolution of Echo 30 has occurred in China .

Miocarditis virales como causa de muerte súbita, aproximación al diagnóstico molecular en patología forense / Viral myocarditis as a cause of sudden death, molecular diagnostic approach to forensic pathology

En Costa Rica, como en el resto del hemisferio occidental, las muertes súbitas tienen principalmente un origen cardiaco. Dentro de este grupo predominan abrumadoramente las que se originan por cardiopatía isquémica, sin embargo no hay que dejar de lado el grupo de las miocarditis. Estas enfermedades la mayor parte de las veces pasan desapercibidas, pero en ocasiones debutan con muerte súbita y es aquí donde cobran importancia médico legal. En el presente artículo se hace una revisión de las miocarditis, con énfasis en las de origen viral, que morfológicamente puede sospecharse su diagnóstico, pero la confirmación del mismo debe ser molecular, siendo una de las indicaciones de este tipo de estudios en patología forense.

In Costa Rica, as in the rest of the western hemisphere, sudden deaths are mainly of cardiac origin. Within this group overwhelmingly dominate those originating from ischemic heart disease; however there is no need to set aside the group of myocarditis. These diseases most often go unnoticed, but sometimes present with sudden death and this is where forensic become important. This article is a review of the myocarditis, with emphasis on the viral origin which morphologically diagnosis may be suspected, but confirmation of the same should be molecular, one of the indications of this type of study in forensic pathology.

Study on the genetic characteristics and molecular epidemiology of Echovirus 11 strains isolated from patients with hand food and mouth disease in He-nan province, China / 中华微生物学和免疫学杂志

Objective To analyze the evolution and genetic characteristics of Echovirus 11 (Echo11) strains isolated from patients with hand food and mouth disease ( HFMD) in Henan province. Methods Enterovirus strains were isolated from stool samples of patients with HFMD from year 2010 to 2012.The sequences of VP1 region of all positive strains were amplified , sequenced and then analyzed by BLSAT to determine their genotypes .The entire VP1 coding regions of 10 Echo11 strains were amplified by RT-PCR, and the homology analysis was conducted among them and other Echo 11 strains published in Gen-Bank.A phylogenetic tree was constructed to evaluate the evolution of Henan isolates and the prevalence of different genotypes .Results From year 2010 to 2012 , 184 non-Enterovirus 71 ( EV71 ) strains and non-Coxsakievirus A16 (CA16) strains causing HFMD were isolated, 10 (5.43%) of with were Echo11 strains. There were 876 nucleotides ( nt) in the complete VP1 gene sequence , encoding 292 amino acids ( aa) .The 10 Echo11 strains shared 93.1%-100% homologies in nt sequences and 97.3%-100% in aa sequences. The homology analysis indicated that Henan strains and prototype strains were highly similar but different with the homology of 77.8%-78.8% in nt and 90.8%-91.8% in aa, respectively.Conclusion Echo11 was one of the pathogens causing HFMD in He-nan province and genotype A was the dominant genotype .

Complete nucleotide sequence of a human echovirus 6 strain KM57-09 isolated in Yunnan,China,in 2009 / 中华流行病学杂志

Objective To analyze the genetic characterization of the complete genome from a human echovirus 6 (Echo6) strain KM57-09 isolated in Yunnan,China,in 2009.Methods Using the RT-PCR,eight fragments containing about 1000 nucleotides which covered the whole viral genome were sequenced.The sequences were aligned with other reference cnterovirus sequences downloaded from the GenBank,using Mega 5.05,RDP 3 and SimPlot 3.5.1 softwares.Results Similar to the other human cntcrovirus,KM57-09 isolate genome appeared to have 7419 nucleotides in length,encoding for 2191 amino acids.In the complete genome,the rates of homology on nucleotide and amino acid among the seven Echo6 isolates were 79.3％-80.2％ and 93.3％ 94.4％,respectively as well as 79.3％ and 93.6％ of the rates of homology when compared with that of D' Amor prototype strain.In different segment of genome.The 2C 3A genome region was most similar to the HN-2-E25 strain,the 5' UTR,VP4,3D and 3' UTR genomc region were most similar to the CoxB5-Henan2010.In the VP1 gene,the rates of homology on nuclcotide and amino acid among the China isolates were 80.0％-96.0％ and 95.8％-99.0％,respectively,and showed 77.6％-96.0％ and 95.2％-99.0％ of the rates on homology when compared to the other Echo6 reference strains isolated from other countries or areas,respectively.Results from phylogenetic analysis showed that the Echo6 formed five distinct groups,A-E.The KM57-09 strain belonged to clade E.The nucleotide divergence between clades was 15.6％-23.3％.The putative recombinant event for KM57-09 was detected with RDP 3,SimPlot 3.5.1 and 3D sequence phylogenetic analysis.Conclusion All the Echo6 isolates could be divided into five clades,the KM57-09 strain belonged to Clade E.The Echo6 strains isolated in China were contributed to several different chains of transmission.

A viral meningitis outbreak associated with Echo30 in drinking water / 中华流行病学杂志

Objective To study the source of infection,the scope of epidemic and control measures in an outbreak involving students having symptoms as fever,dizziness,headache,vomiting and nausea.Methods The suspected-case was defined as fever (armpit temperature ≥37 ℃) and with one or more of the following symptoms:dizziness,headache,vomiting and nausea,among students and teachers at school from Mar 1,2012.Confirmed-case was among suspected case accompanied by both throat and rectal swabs enterovirus positive by RT-PCR.All the cases were collected through checking the medical records from 4 hospitals as well as through the absence records of students and teachers,from Mar 1,2012.We conducted a case-control study with ratio of 1 ∶ 2 and data on the exposures to water among students and teachers was collected prior to the illness.27 cases' throat and rectal swabs were collected and analyzed by RT-PCR and PCR sequence methods.2 warm-water samples were collected for testing the counts on total bacteria and E.coli.Results 103 students' cases were identified in school L,with the attack rate as 4.6％ (103/2255).Students from Grade three had the high attack rate as 18.1％ (72/397) and 77.7％ (80/103) of the cases located in the building with ' multiple-functions'.Epidemic curve of the outbreak showed a pattern with continuous common source of infection.It seemed that the exposure to warm-water appeared to be the major risk factor (OR =18.3,95％CI:2.0-169.5) together with the intake of un-boiled water (OR =15.5,95 ％CI:1.7-141.8).Specimens from 27 students (81.5％,22/27) were identified enterovirus positive by RT-PCR,and 7 of the 9 students were confirmed carrying Echo 30.Bacteria and coli were negative from the 2 warm-water samples.Conclusion This viral meningitis-outbreak was caused by Echo 30,with drinking water as the major risk factor.

Molecular epidemiology of human Parechovirus infections in central nervous system-related disease and spsis in children / 中华传染病杂志

Objective To obtain the molecular epidemiology of human Parechovirus (HPeV)infections m children with central nervous system (CNS)-related disease and sepsis,as well as understand the pathogenic properties of HPeV infections by detecting HPeV in cerebrospinal fluid (CSF) and blood samples.Methods From January to December in the year of 2009,a total of 359enterovirus-negative specimens including 210 CSF and 149 blood samples were collected from 328children ＜14 years of age who were hospitalized for CNS-related disease and sepsis at Children's Hospital,Fudan University,Shanghai,China.HPeV was detected by nested reverse transcription polymerase chain reaction (RT-PCR),and then directly genotyped by sequencing nested RT-PCR product of VP3/VP1 region.Ninty-nine blood samples from healthy children were collected as controls during the same period.Results Twenty-seven children (8.2％) were HPeV positive in 328 children.HPeV infections were found in all age groups of children and the highest frequency was seen in children ＜3 months old (18.2％,12/66).HPeV was detected in several months,with the peak in December (18.8％,9/48).Of all the positive samples,20 were genotyped successfully and identified to be HPeV1.No HPeV infections were found in blood of healthy controls.ConclusionsHPeV is the pathogen of CNS infections and sepsis in children.HPeV screening should be enrolled in the routine virus testing in specimens obtained from children.HPeV1 is the prevalent type in children in the year of 2009 in Shanghai.