A coxsackievirus B5‐associated aseptic meningitis outbreak in Shandong Province,China in 2009

In 2009, a major outbreak of aseptic meningitis was noted in Linyi city, Shandong province, China. From June to September 2009, a total of 2,104 cases were involved in this outbreak, and 98.6% of patients were <16 years of age. To determine the pathogen of the outbreak, 42 cerebrospinal fluid specimens collected from aseptic meningitis cases were tested for cell culture, and 17 (40.5%) enteroviruses were isolated and identified as Coxsackievirus B5 (CVB5). Homologous comparison indicated that these isolates had 0–7.7% nucleotide divergence with each other. Phylogenetic reconstruction showed global CVB5 could be separated into four genogroups, and all Linyi CVB5 isolates belonged to the genogroup C which had been circulating for recent 27 years in Asia and Europe. Interestingly, two distinct lineages were observed for the 17 isolates in the phylogenetic tree, indicating that at least two different transmission chains of CVB5 were responsible for this outbreak. This study showed that CVB5‐associated aseptic meningitis is an emerging concern in China. J. Med. Virol. 85:483–489, 2013. © 2012 Wiley Periodicals, Inc.     

RT-nPCR Assays for Amplification and Sequencing of VP1 Genes in Human Enterovirus A-D from Clinical Specimens

ObjectiveTo develop RT-nPCR assays for amplifying partial and complete VP1 genes of human enteroviruses (HEVs) from clinical samples and to contribute to etiological surveillance of HEV-related diseases.MethodsA panel of RT-nPCR assays, consisting of published combined primer pairs for VP1 genes of HEV A–C and in-house designed primers for HEV-D, was established in this study. The sensitivity of each RT-nPCR assay was evaluated with serially diluted virus stocks of five serotypes expressed as CCID₅₀ per μL and copies per μL, and the newly established methods were tested in clinical specimens collected in recent years.ResultsThe sensitivity of RT-nPCR assays for amplifying partial VP1 gene of HEVs was 0.1 CCID₅₀ per μL and 10 virus copies per μL, and for the complete VP1 gene was 1 CCID₅₀ per μL and 100 virus copies per μL, using serially-diluted virus stocks of five serotypes. As a proof-of-concept, 25 serotypes were identified and complete VP1 sequences of 23 serotypes were obtained by this system among 858 clinical specimens positive for HEVs during the past eight surveillance seasons.ConclusionThis RT-nPCR system is capable of amplifying the partial and complete VP1 gene of HEV A–D, providing rapid, sensitive, and reliable options for molecular typing and molecular epidemiology of HEVs in clinical specimens.     

引起儿童呼吸道感染的优势肠道病毒及其血清型和流行特征分析

目的 了解近年杭州地区引起儿童呼吸道感染的优势肠道病毒(EVs)及其血清型以及流行特征和易感人群。方法 收集2016-2018年本地区2 164例发热伴呼吸道感染症状患儿咽拭标本,采用EVs通用引物的实时荧光定量RT-PCR检测EVs核酸,阳性标本采用EVs VP1/VP2基因通用引物的套式PCR扩增后测序分型。分析患儿性别、年龄、感染的优势EVs及其血清型等流行病学特征。结果 2 164 例患儿咽拭子标本中有508 例(23.5%)EVs核酸阳性,其中290 例成功测序分型。检出的EVs中,柯萨奇病毒A 组(CoxA)和B 组(CoxB)分别占75.2%和11.7%、EV71占8.9%、埃可病毒(Echo)占4.1%,CoxA检出率高于CoxB、EV71和埃可病毒(P<0.05)。在检出的CoxA中,CoxA6(39.5%)和CoxA10(33.0%)检出率高于CoxA16、CoxA4和CoxA2(P<0.05)。1~3 岁男性儿童是EVs易感人群(P<0.05),其中男性儿童易感CoxA6、女性儿童易感CoxA10(P<0.05)。结论 EVs是本地区儿童呼吸道感染的重要病原体,柯萨奇病毒及其CoxA6和CoxA10分别是优势EVs和血清型,1~3 岁儿童是本地区EVs主要易感人群,男性比女性更易感。     

High Heterogeneity of Echoviruses in Brazilian Children with Acute Gastroenteritis

Echoviruses (E) are a diverse group of viruses responsible for various pathological conditions in humans including aseptic meningitis, myocarditis, and acute flaccid paralysis. The detection and identification of echovirus genotypes in clinical samples is challenging due to its high genetic diversity. Here, we report the complete genome sequences of nine echoviruses, obtained by next-generation sequencing of 238 fecal samples from individuals with gastroenteritis in regions of Brazil. Detected viruses were classified into six genotypes: Three E1 sequences (BRA/TO-028, BRA/TO-069 and BRA/TO-236), one E3 (BRA/TO-018), one E11 (BRA/TO-086), one E20 (BRA/TO-016), two E29 (BRA/TO-030 and BRA/TO-193), and one E30 sequence (BRA/TO-032). Phylogenetic analysis indicated that the echoviruses E1 and E29 circulating in Brazil are divergent from strains circulating worldwide. The genotype diversity identified in our study may under-represent the total echovirus diversity in Brazil because of the small sample size and the restricted geographical distribution covered by the survey.     

Circulation and Molecular Epidemiology of Enteroviruses in Paralyzed,Immunodeficient and Healthy Individuals in Tunisia,a Country with a Polio-Free Status for Decades

This report is an overview of enterovirus (EV) detection in Tunisian polio-suspected paralytic cases (acute flaccid paralysis (AFP) cases), healthy contacts and patients with primary immunodeficiencies (PID) during an 11-year period. A total of 2735 clinical samples were analyzed for EV isolation and type identification, according to the recommended protocols of the World Health Organization. Three poliovirus (PV) serotypes and 28 different nonpolio enteroviruses (NPEVs) were detected. The NPEV detection rate was 4.3%, 2.8% and 12.4% in AFP cases, healthy contacts and PID patients, respectively. The predominant species was EV-B, and the circulation of viruses from species EV-A was noted since 2011. All PVs detected were of Sabin origin. The PV detection rate was higher in PID patients compared to AFP cases and contacts (6.8%, 1.5% and 1.3% respectively). PV2 was not detected since 2015. Using nucleotide sequencing of the entire VP1 region, 61 strains were characterized as Sabin-like. Among them, six strains of types 1 and 3 PV were identified as pre-vaccine-derived polioviruses (VDPVs). Five type 2 PV, four strains belonging to type 1 PV and two strains belonging to type 3 PV, were classified as iVDPVs. The data presented provide a comprehensive picture of EVs circulating in Tunisia over an 11-year period, reveal changes in their epidemiology as compared to previous studies and highlight the need to set up a warning system to avoid unnoticed PVs.     

泰安市2012-2013年托幼机构手足口病感染状况分析

目的对2012-2013年度泰安市托幼机构HFMD病例的标本进行病原体检测和分析,了解本地区托幼机构HFMD感染状况。方法对临床诊断为手足口病病例在园儿童的粪便标本249份,用实时荧光定量PCR检测肠道病毒EV71和CA16。结果 2012-2013年肠道病毒阳性率分别为83.33%(110/132)和93.16%(109/117)。肠道病毒总阳性率为87.95%(219/249),CA16阳性率为40.56%(101/249),EV71阳性率为28.92%(72/249),其他肠道病毒为18.47%(46/249),CA16的阳性率高于EV71。发病年龄主要集中在2~4岁,男女性别比例为1.47:1。结论泰安市托幼机构手足口病疫情存在明显的年龄、性别差异,多种病毒株共存,以CA16为主。     

贵州省从江县81例手足口病病例病原学检测结果

目的了解贵州省从江县2011—2012年手足口病流行的主要病原体,为手足口病防控提供科学依据。方法采集临床诊断为手足口病病例的咽拭子采用RT-PCR方法扩增病毒的特异性片段,检测EV71、CoxA16核酸和其他肠道病毒核酸。结果共采集标本81份,病毒核酸检测阳性33份,阳性率为40.74%;其中EV71阳性9份（27.27%）,CoxA16阳性4份（12.12%）,其他肠道病毒阳性20份（60.61%）。结论从江县2011—2012年手足口病流行毒株以其他肠道病毒为主。但也不能忽视EV71引起的有中枢神经系统并发症的重症病例的监测。