惠州市重症手足口病病原谱及EV71型VP1区基因特征分析

目的分析惠州市手足口病重症病例的病原谱构成,了解惠州市肠道病毒71型分离株的VPl区基因特征．为科学防治手足口病提供科学依据。方法采集300例重症手足口病（HFMD）患者标本,采用实时荧光RT-PCR检测肠道病毒核酸,并对肠道病毒7l型（EV71）和柯萨奇病毒A16型（CoxA16）进行分型检测;选择8株EV71分离株进行VPl区基因全长序列测定,测序结果利用DNASTAR软件进行核苷酸、氨基酸序列分析和同源性比较。并用Mega5．0软件构建亲缘性进化树。结果通过实时荧光RT-PCR特异性检测,EV71阳性结果154份,阳性率为51．33％;CoxAl6阳性结果38份,阳性率为12．67％。测序结果表明,8株EV71之间的VPl基因核苷酸同源性为96．9％～99．2％。氨基酸同源性为99．3％～100％。VPI区基因遗传进化分析表明。8株EV71分离株与c4基因亚型的代表株处于同一分支,均属于C4基因亚型的C4a进化分支。结论惠州市手足口病疫情主要病原EV71病毒均属于C4a基因亚型,与2004年以来的中国大陆EV71病毒流行的基因型一致,未产生明显的抗原漂移及变异。     

2009年夏季174例手足口病患儿病原分析

目的分析本院2009年夏季手足口病患儿病原及其血清型特征,为临床早期诊断和疾病防控提供实验依据。方法2009年4—9月,首都儿科研究所附属儿童医院门诊就诊的174例手足口病患儿咽拭子和疱疹液,实时荧光RT—PCR方法检测肠道病毒通用型（EV）、柯萨奇A16型（CA16）、肠道病毒71型（EV71）。131例患儿双份血清,同时检测CA16、EV71IgM抗体。CA16和EV71阳性标本扩增VP1区基因片段后测序,进行同源性和系统进化分析。结果（1）EV、CA16、EV71阳性例数分别为167、112、46;阳性率为96．O％、64．4％、26．4％,CA16：EV71为2．43：1。（2）首诊血清CA16和EV71IgM阳性例数为51、25,阳性率为38．9％、19．1％,复诊阳性例数为98、32,阳性率为74．8％、24．4％。（3）CA16VP1区核苷酸同源性为88．7％-98．5％,EV71VP1区核苷酸同源性为94．9％-99．7％,与c4亚型参比序列核苷酸同源性92．1％～95．3％。结论2009年夏季本院手足口病患儿病原以CA16、EV71为主,EV71阳性率较之前报道有较大幅度升高。EV71病毒株以c4亚型为主。实时RT-PCR法较血清学检测特异性IgM抗体更适于疾病早期诊断。     

2010—2018年乐清地区肠道病毒感染疾病的病原特征分析

目的分析乐清地区2010—2018年引起儿童肠道病毒(EV)相关疾病的病原谱变化及其各型别的流行趋势,为肠道病毒引起手足口病防控工作提供科学依据。方法从乐清市疾病监测信息报告管理系统导出2010—2018年肠道病毒感染引起手足口病病例资料,采用描述性流行病学方法分析手足口病病例发病的季节、年龄、性别、区域分布及病原体的分布特点。结果2010—2018年乐清市手足口病病例共计53178例。全年各月均有发病,发病高峰在4—7月,重症75例,死亡6例,5岁以下的散居儿童和托幼儿童发病为主,男性高于女性。2010—2018乐清地区儿童中感染的病原谱发生了明显变化,2010—2014年均以EV71型为主,2013年之后EV71检出比例逐年降低,近几年来非EV71非柯萨奇病毒A16型的其他肠道病毒比例明显升高。结论乐清地区肠道病毒引起手足口病具有明显的季节性、人群性,应加强病原学检测,在4—7月,对重点对象采取综合防控措施,防止引起手足口病暴发流行。     

黄石市2012年手足口病的流行特征及病原学分析

目的了解黄石市2012年手足口病的流行病学特征及规律,为科学制定手足口病的防控策略提供依据。方法利用国家疾病监测信息管理系统获得2012年手足口病疫情资料．对2012年1—12月8957例手足口病病例资料进行统计分析。结果黄石地区手足口病的流行从3月份开始增多,4—5月份急剧上升至高峰,随后逐月下降,11-12月病例数又出现轻微的抬升。发病集中在4—6月份,发病病例占全年病例数的2／3。3岁以下发病人群中男女比例为1．91：1．男性显著性高于女性（X^2=4．29,P〈0．05）,且70％以上为散居儿童。病原学分析显示,2012年以肠道病毒71型为主导。结论手足口病的发生有明显的季节、年龄、性别和职业差异,掌握手足口病的流行病学特征,将有助于更好地开展预防和控制工作。     

临沂地区1426例手足口病流行病学及临床特征分析

目的探讨山东省临沂地区手足口病的流行病学及临床特征,为制定本地区疾病防治措施提供依据。方法制定手足口病观察项目表,采集2012年4—9月山东省临沂地区1426例手足口病患者的流行病学及临床特征,果用描述流行病学方法进行分析。结果临沂地区手足口病发病高峰为5月,以3岁以下散居及幼托儿童为主,发病率男童多于女童,农村高于城镇。临床表现主要包括发热、皮疹等,重症患儿可发生多系统并发症。辅助检查可见白细胞及中性粒细胞比值增高等特征。EV71为重症病例的主要病原,占重症病例的77．21％。结论山东省临沂地区手足口病流行具有明显季节性、人群性及地区性。针对本地区流行病特点,制定区域特异性防治措施至关重要。     

Enterovirus spectrum from the active surveillance of hand foot and mouth disease patients under the clinical trial of inactivated Enterovirus A71 vaccine in Jiangsu,China, 2012‐2013

Epidemiological data from active surveillance on human enterovirus, which could cause hand, foot, and mouth disease, were limited. An active surveillance system was used to investigate the enterovirus spectrum and the incidence of different enteroviruses in infants aged 6–35 months in Jiangsu Province from 2012 to 2013. Fifty‐nine infants were randomly selected from 522 non‐EV‐A71/CV‐A16 HFMD patients. We collected 173 throat swabs and 174 rectal swabs from these infants. RT‐PCR was used to amplify 5'‐UTR and VP1 regions of enteroviruses and the serotypes were determined by the sequence comparison using BLAST. Twenty‐one non‐EV‐A71/CA16 enterovirus serotypes were detected in those infants. E16, E18 were firstly reported in HFMD patients. The four top common non‐EV‐A71/CV‐A enteroviruses among infants were CV‐B3, CV‐A10, CV‐A6, and E9 with the HFMD incidence rates at 1.4%, 0.84%, 0.56%, and 0.47%, respectively. Over 20.8% patients were co‐infected with multiple enteroviruses. Neither the course of sickness nor clinical symptoms of the co‐infected patients was more severe than those infected with single enterovirus. Two patients were infected different enterovirus successively within 2 months. Several new enterovirus serotypes and multiple models of infection associated with HFMD were discovered through the active surveillance system. These data provide a better understanding of the viral etiology of HFMD. J. Med. Virol. 87:2009–2017, 2015. © 2015 Wiley Periodicals, Inc.

     

东莞市2009-2010年手足口病疫情分析及防治对策探讨

目的了解东莞市2009-2010年手足口病疫情的流行病学特点,为今后的防控工作提出对策。方法对东莞市2009-2010年手足口病疫情资料采用描述流行病学方法进行分析,对患者的咽拭子或粪便标本采用RT-PCR法进行肠道病毒核酸检测和病毒分型。结果东莞市2009年和2010年分别报告手足口病9263例和22721例,发病率分别为127.34/10万和373.28/10万。5岁以下婴幼儿分别占当年报告病例的91.68%和92.15%;散居儿童分别占当年报告病例的78.26%和73.58%;男性分别占当年报告病例的65.29%和65.27%;每年的4月份和10月份有2个发病高峰,前者为全年的发病高峰;暴发疫情均发生在托幼机构;分别有49.30%和53.65%的病例由EV71引起,所有死亡病例均由EV71引起。健康人群肠道病毒隐性感染率为11.67%,感染的主要病原体为其他肠道病毒（95.24%）。结论东莞市2010年手足口病流行强度大于2009年;应重点做好手足口病的病原学监测及5岁以下散居儿童和托幼机构的手足口病防治工作。     

实时RT-PCR检测159例手足口病患儿样本中肠道病毒的结果分析

目的 了解2009年4-8月首都儿科研究所附属儿童医院手足口病患儿肠道病毒的感染状况,为临床诊治提供参考.方法 采集首诊手足口病159例患儿的咽拭子和疱疹液标本,以肠道病毒(EV)通用型、柯萨奇病毒A16(CA16)型、肠道病毒71(EV71)型核酸检测试剂盒,应用实时RT-PCR法检测标本中的肠道病毒.选取阳性标本扩增VP1区,产物进行序列测定和分析.结果 (1)EV、CA16、EV71的阳性病例数分别为152、102、43;阳性率为95.6%、64.2%、27.0%.(2)CA16占EV阳性的67.3%,EV71占EV阳性的28.3%,非CA16和EV71的EV病例7例,占EV阳性的4.6%.CA16:EV71为2.37:1.(3)部分阳性标本经测序验证与此法结果一致.结论 2009年我院手足口病患儿以EV71和CA16感染为主,EV71感染的手足口病比例较2007年出现明显上升.     

Diagnosis of hand,foot, and mouth disease caused by EV71 and other enteroviruses by a one‐step,single tube,duplex RT‐PCR

Hand, foot, and mouth disease (HFMD) is caused mainly by enterovirus 71 (EV71) and other enteroviruses (EVs) such as Coxsackie A16 in China. EV71 infection can lead to severe clinical manifestations and even death. Other EVs, however, generally cause mild symptoms. Thus, early and accurate distinction of EV71 from other EVs for HFMD will offer significant benefits. A one‐step, single tube, duplex RT‐PCR assay is described in the present study to detect simultaneously EV71 and other EVs. The primers used for the duplex RT‐PCR underwent screening and optimization. The detection threshold was 0.001 TCID₅₀/ml for EV71 and 0.01 TCID₅₀/ml for other EVs. The positive rate of enterovirus detection in 165 clinical samples reached 68.5%, including 46.1% for EV71 and 22.4% for other EVs. Of all the severe HFMD cases, EV71 was responsible for 85.3% cases. The positive rate of EV71 fell markedly by day 8 after onset. In addition, sequencing of EV71 specific amplicons from duplex RT‐PCR revealed that C4a was the predominant subgenotype of EV71 circulating in Nanjing, China. The accuracy and reliability of the assay suggest strongly that the one‐step, single tube, duplex RT‐PCR will be useful for early diagnosis and monitoring of EV71 and other EV infections. J. Med. Virol. 84:1803–1808, 2012. © 2012 Wiley Periodicals, Inc.     

湖北省宜昌市2008—2012年手足口病流行病学特征分析

目的分析湖北省宜昌市2008—2012年手足口病疫情流行特征,为制定预防控制措施提供科学依据。方法采用描述性流行病学方法,对宜昌市2008—2012年手足口病发病情况和病原检测情况进行分析。结果2008—2012年宜昌市报告手足口病20774例,年均报告发病率为104．80／10万,重症2例,无死亡病例报告。主要集中在0~6岁儿童,占总病例数的92．19％（19152／20774）。男性发病率为121．53／10万,高于女性的86．99／10万,差异有统计学意义（X2=564．423,P〈0．01）。宜昌市5个市辖区年均报告发病率为134．57／10万,高于其余8个农村县市的89．83／10万,差异有统计学意义（x2=843．08,P〈0．01）。职业分布以散居儿童（46．99％）和幼托儿童（46．62％）为主;发病高峰为4—7月,次高峰11—12月。实验室确诊占报告总病例的3．32％（689／20774）,样本检测阳性率60．12％（689／1146）。确诊病例中,EV71型（220例）占31．93％,CoxAl6型（271例）占39．33％,其他肠道病毒（198例）占28．74％。结论宜昌市手足口病发病有明显的季节、地区、年龄、性别差异,发病逐年上升。应加强疫情监测和宣传教育力度,有针对性落实重点季节、重点地区、重点人群、重点环节防控措施是防控手足口疫情的关键。     

Relationship between serologic response and clinical symptoms in children with enterovirus 71-infected hand-foot-mouth disease

This study aimed to explore the correlation between clinical symptoms, including rash and fever, and serum antibody reaction to enterovirus 71 (EV71) infection in children hospitalized due to hand-foot-mouth disease (HFMD). From May 2014 to July 2014, a total of 547 children hospitalized due to HFMD in Children’s Hospital of Fudan University were enrolled retrospectively. RNA levels of EV71 and CA16 in fecal, serum, and cerebrospinal fluid specimens were measured using quantitative real-time RT-PCR, and EV71-IgM antibody in the serum was detected using immune colloidal gold assays. Of the 547 fecal specimens, 296 were EV71 RNA positive, 109 were CA16 RNA positive, and 8 were positive for both EV71 RNA and CA16 RNA. The total positive rate for either EV71 or CA16 in feces was 72.58% (397/547). Additionally, 544 serum specimens were collected, and 409 were EV71-IgM positive (75.18%). The duration of rash and fever was found to be correlated to the positive rate of serum EV71-IgM, and the positive rate of serum EV71-IgM plus EV71 RNA in feces. The positive rates of serum EV71-IgM and serum EV71-IgM plus EV71 RNA in fecal collected at day 3 of fever were 79.7% and 52.8%, respectively. In conclusion, EV71 and CA16 were found to be the major pathogens responsible for the epidemics of HFMD in children during May to July 2014 in Shanghai, China. There is a close relationship between the positive rate of serum EV71-IgM and the duration of fever and rash.     

Coxsackievirus A6 and enterovirus 71 causing hand,foot and mouth disease in Cuba, 2011–2013

Hand, foot and mouth disease (HFMD) is usually caused by coxsackievirus A16 or enterovirus 71 (EV71). Between 2011 and 2013, HFMD cases were reported from different Cuban provinces. A total of 42 clinical specimens were obtained from 23 patients. Detection, identification and phylogenetic analysis of enterovirus-associated HFMD were carried out by virus isolation, specific enterovirus PCR and partial VP1 sequences. HEV was detected in 11 HFMD cases. Emerging genetic variants of coxsackievirus A6 and EV71 were identified as the causative agents of the Cuban HFMD cases.     

An outbreak of enterovirus 71 infection in Taiwan, 1998: epidemiologic and clinical manifestations.

BACKGROUND: An outbreak of enterovirus infections occurred throughout Taiwan in 1998. The diseases were manifectated with hand, foot, and mouth disease (HFMD), some associated with meningitis, encephalitis, or acute flaccid paralysis (AFP). OBJECTIVES: This study is aimed to characterize and analyze the epidermologic and clinical features during the outbreak. STUDY DESIGN: The epidemiologic information was collected from the Ministry of Health on passive surveillance; clinical and virological investigations were carried out at National Cheng Kung University Medical Center. RESULTS: Between April and December 1998, 405 children were hospitalized, and 78 patients died during this outbreak in Taiwan. There were 119 cases identified to be EV71 infection in Tainan and Chiayi areas; 105 cases by virus isolation and 14 by serological assay. The outbreak had a biphasic curve with peak in June and October, especially in the southern Taiwan. Seventy-two percent of patients were below 3 years of age. The spectrum of disease included HFMD in 54, HFMD with central nerve system (CNS) involvement in 37, herpangina in 12, aseptic meningitis in three, encephalitis/ meningoencephalitis in ten, acute flaccid paralysis in three. There was nine fatal cases complicated with neurogenic pulmonary edema. Myoclonus with sleep disturbance was the most important early sign of EV71 infection with CNS involvement. CONCLUSION: Our experience demonstrated that the EV71 isolated in Taiwan had strong dermatotropic as well as neurotropic tendencies. Early detecting CNS involvement and commencing aggressive therapy may reduce the mortality.     

深圳市重症手足口病临床与实验特征变化趋势及随访1年报告

目的比较2012与2008年深圳市重症手足口病临床和实验特征,探讨其变化趋势。方法深圳市第三人民医院2008年及2012年共收治778例手足口病住院患儿,重点分析其中260例手足口病重症患者（2008年64例,2012年196例）。通过RT—PCR检测CoxsackieA16（CoxA16,A16）及Enterovirus71（EV71）病毒核酸,出现神经系统症状或体征者检查脑脊液常规及生化,部分患儿行头颅MRI检查,同时比较临床表现、血液生化等变化趋势。所有患者随访12个月评估再发情况。结果与2008年相比,2012年手足口病重症患者发病高峰年龄下降,〈12月龄儿童分别占2008年和2012年重症的15％和40％。两个年度重症患者均以EV71感染为主;与2008年相比,2012年EV71感染所占比率有下降趋势;重症患者手、足皮疹出疹率下降,而口腔疱疹出疹率明显增高,疱疹性咽峡炎引起的重症病例数上升;外周血白细胞计数、中性粒细胞比值及CRP水平均明显增高;但并发脑干脑炎或神经源性肺水肿的病例数略有下降。随访1年结果显示,无论EV71或CoxA16型手足口病临床治愈后均可多次再发,轻症再发率高于重症。结论深圳市手足口病从2008年暴发至2012年,发病呈上升趋势,重症手足口病病例临床表现具有一些新的特征,应引起广大医护人员重视。     

Genetic polymorphism of CCL2-2510 and susceptibility to enterovirus 71 encephalitis in a Chinese population

The study was performed in 36 Chinese patients with enterovirus 71 (EV71) encephalitis and 141 patients with EV71-related hand, foot and mouth disease (HFMD) without encephalitis. Genotyping was done by the polymerase chain reaction-restriction fragment length polymorphism technique. Patients with EV71 encephalitis had a significantly higher frequency of the CCL2-2510GG genotypes when compared to patients with EV71-related HFMD without encephalitis (66.7 % vs. 41.8 %, p = 0.028). The frequency of CCL2-2510G alleles was also significantly higher among the patients with EV71 encephalitis than among patients with EV71-related HFMD without encephalitis (79.2 % vs. 64.9 %, OR = 2.1, 95 % CI = 1.1-3.8, P = 0.023). Significant differences were found in gender, age, fever days, white blood cell count, C-reactive protein level, blood glucose concentration, and CCL2 level among genotypes of CCL2-2510A/G in EV71-infected patients, but no significant differences were found in alanine aminotransferase, aspartate aminotransferase, or creatine kinase myocardial isozyme levels or in cerebrospinal fluid evaluations (except monocytes) in patients with EV71 encephalitis. These findings suggest that the CCL2-2510G allele is associated with susceptibility to EV71 encephalitis in Chinese patients.     

成都市龙泉驿区2011-2015年手足口病流行特征分析

目的 对四川省成都市龙泉驿区2011-2015年手足口病流行特征进行分析,为今后制定和修改防控策略提供依据.方法 对龙泉驿区2011-2015年手足口病监测资料进行描述性统计分析及时间和空间扫描统计分析.结果 龙泉驿区5年间共报告手足口病病例9 067例,年平均发病率210.83/105;重症病例241例,占病例总数的2.66％,死亡2例.1-3岁散居儿童为该病的高发人群.4-7月和10-12月是发病的高峰月份.EV71是重症病例和死亡病例的主要病原体.聚集性疫情主要集中于幼托机构,市级幼儿园聚集性疫情发生最少.结论 2011-2015年龙泉驿区手足口病发病水平呈逐年增高趋势,应重点加强散居儿童的社区防控力度,落实幼托机构的防控措施.     

2007-2015年北京市密云区手足口病流行特征分析

目的 分析2007-2015年密云区手足口病流行病学特征,为调整防控措施提供科学依据.方法 采用描述性流行病学方法,对密云区2007-2015年手足口病监测数据进行分析.结果 2007-2015年密云区手足口病报告发病数及发病率每2-3年有一发病高峰,发病时间以4-9月份为主;手足口病高发地区主要为城区及城乡结合部的平原地区;患者以5岁以下儿童为主;男童病例数高于女童;EV71和CVA16为手足口病的主要致病病原体.结论 密云区手足口病报告发病数及发病率呈周期性波动趋势,幼托儿童和散居儿童为手足口病高发人群,城区及城乡结合部的平原地区是手足口病防控的重点区域.     

亳州市2009-2013年手足口病流行病学分析

目的分析亳州市手足口病流行特征,为防控工作提供依据。方法根据中国疾病预防控制信息系统中手足口病发病统计数据,采用描述流行病学方法对亳州市2009-2013年手足口病发病资料进行分析。结果 2009-2013年亳州市共报告手足口病27 088例,年发病率在69.79/10万至109.36/10万之间;全年各月份均有病例报告,发病主要集中在3-6月份（54.60%）;性别比为1.84∶1,1~3岁年龄组占发病总数的80.49%,以散居儿童为主,占95.72%,病原学检测EV71病毒占70.67%。结论亳州市手足口病疫情3-6月份为发病高峰,病原学以EV71病毒为主,加强监测、宣传教育是减少手足口发病的重要措施。     

Molecular epidemiology of enterovirus 71, coxsackievirus A16 and A6 associated with hand,foot and mouth disease in Spain

Hand, foot and mouth disease (HFMD) is a childhood illness frequently caused by genotypes belonging to the enterovirus A species, including coxsackievirus (CV)-A16 and enterovirus (EV)-71. Between 2010 and 2012, several outbreaks and sporadic cases of HFMD occurred in different regions of Spain. The objective of the present study was to describe the enterovirus epidemiology associated with HFMD in the country. A total of 80 patients with HFMD or atypical rash were included. Detection and typing of the enteroviruses were performed directly in clinical samples using molecular methods. Enteroviruses were detected in 53 of the patients (66%). CV-A6 was the most frequent genotype, followed by CV-A16 and EV-71, but other minority types were also identified. Interestingly, during almost all of 2010, CV-A16 was the only causative agent of HFMD but by the end of the year and during 2011, CV-A6 became predominant, while CV-A16 was not detected. In 2012, however, both CV-A6 and CV-A16 circulated. EV-71 was associated with HFMD symptoms only in three cases during 2012. All Spanish CV-A6 sequences segregated into one major genetic cluster together with other European and Asian strains isolated between 2008 and 2011, most forming a particular clade. Spanish EV-71 strains belonged to subgenogroup C2, as did most of the European sequences circulated. In conclusion, the recent increase of HFMD cases in Spain and other European countries has been due to a larger incidence of circulating species A enteroviruses, mainly CV-A6 and CV-A16, and the emergence of new genetic variants of these viruses.