肠道病毒71型和柯萨奇病毒A16型细胞受体的研究进展

近几年手足口病在亚太地区的流行大幅上升并伴随严重的中枢神经系统并发症,而针对其主要致病原肠道病毒71型和柯萨奇病毒A16型现在还缺乏有效的预防疫苗和抗病毒制剂.确定它们的特异性细胞受体对于进一步阐明病毒与宿主细胞早期互相作用的分子机制及其致病机制至关重要.过去一年中,P-选择素糖蛋白配体-1、B类清道夫受体Ⅱ和唾液酸化...     

多重荧光逆转录PCR同时检测肠道病毒71型和柯萨奇病毒A16型

目的建立多重荧光逆转录PCR（RT-PCR）同时检测肠道病毒71型和柯萨奇病毒A16型的方法。方法在肠道病毒71型和柯萨奇病毒A16型VP1基因的保守区序列分别设计特异性引物和Taqman探针,建立优化多重荧光RT-PCR反应体系,评价所建多重荧光RT-PCR反应体系的特异性、敏感性和稳定性,并应用于临床样品检测。结果该方法对肠道病毒71型和柯萨奇病毒A16型检测具有高度特异性,检出限分别为0.1 TCID50和1 TCID50,具有较好的稳定性,可直接应用于临床样品的检测。结论本研究建立的多重荧光RT-PCR可以同时准确检测肠道病毒71型和柯萨奇病毒A16型,灵敏度高,稳定性好,是一种快速检测肠道病毒71型和柯萨奇病毒A16型的新方法。     

人类肠道病毒71型研究进展

手足口病(Hand-Foot-Mouth disease,HFMD)是由肠道病毒引起的一种常见疾病,严重危害儿童健康.2008年5月被列入《中华人民共和国传染病防治法》法定丙类传染病.其病原体复杂,引发手足口病的肠道病毒有20多种(型),包括柯萨奇病毒A组的16、4、5、9、10型等,B组的2、5型等,以及肠道病毒71型等,其中以柯萨奇病毒A16型(Cox A16)和肠道病毒71型(EV 71)最为常见.EV71病毒与CA16型病毒感染虽然在临床表现上比较接近,但EV71病毒还可以引起如无菌性脑膜炎、脑干脑炎和脊髓灰质炎样的麻痹等多种中枢神经系统疾病,致死率较高,因此备受关注.     

2011年南京儿童手足口病病原学特征分析

目的分析2011年南京儿童手足口病病原学特征,为手足口病的临床治疗提供科学的参考依据。方法对2011年3—7月流行季节南京地区186例手足口病患儿进行肠道病毒E71型(EV71)、卡萨奇病毒A16型(CoxA16)和其他肠道病毒的监测,监测所得资料采用流行病学方法进行分析。结果在186例手足口病患儿中普通病例以肠道病毒71型和柯萨奇病毒A16型共同主导,重症病例及死亡病例以肠道病毒71型为主要病原体。结论 2011年南京儿童手足口病普通病例中肠道病毒71和柯萨奇病毒A16均可见,重症病例和死亡病例中以肠道病毒71型为主。     

手足口病的口腔表征与专科防护

手足口病是由肠道病毒引起的常见传染病,主要病毒以柯萨奇A16和肠道病毒71型(EV71)为主,<3岁婴幼儿易感.肠道病毒传染性强,易引起暴发流行.     

肠道病毒71型研究进展

手足口病是由不同的肠道病毒所引起的一种急性传染病,其中以柯萨奇病毒A16型和人肠道病毒EV71最为常见。由于EV71能引起严重的中枢神经系统并发症,本文就EV71的研究进展作一综述,为手足口病的预防和治疗提供依据。     

肠道病毒71型研究进展

手足口病是由不同的肠道病毒所引起的一种急性传染病,其中以柯萨奇病毒A16型和人肠道病毒EV71最为常见。由于EV71能引起严重的中枢神经系统并发症,就EV71的研究进展作一综述,为手足口病的预防和治疗提供依据。     

鄞州区2011年手足口病病原学检测结果

<正>手足口病(Hand foot and mouth diease,HFMD)是由肠道病毒引起的一种儿童常见传染病。大多数患者症状轻微,以发热和手、足、口腔等部位的皮疹或疱疹为主要特征。少数患者可并发无菌性脑膜炎、脑炎、急性弛缓性麻痹、呼吸道感染和心肌炎等。该病的病原体主要为柯萨奇病毒A组的16、2、4、5、7、9和11型,柯萨奇病毒B组的1、2、3、4、5和13型,部分埃可病毒和肠道病毒71型。最常见的为柯萨奇病毒A 16型(Cox A16)和肠道病毒71型(EV 71)[1]。     

大仪镇一起幼儿园手足口病疫情调查报告

手足口病是由肠道病毒引起的传染病,多发生于5岁以下儿童,可引起手、足、口腔等部位的疱疹,少数患儿可引起心肌炎、肺水肿、无菌性脑膜脑炎等并发症.个别重症患儿如果病情发展快,导致死亡.引发手足口病的肠道病毒有20多种(型),柯萨奇病毒A组的16、4、5、9、10型,B组的2、5型,以及肠道病毒71型均为手足口病较常见的病原体,其中以柯萨奇病毒A16型(CoxA16)和肠道病毒71型(EV71)最为常见.本文笔者探讨了大仪镇一起幼儿园手足口病疫情调查报告,具体如下.     

肠道病毒71型与手足口病的研究进展

<正>手足口病主要症状表现为手、足、口腔、臂部等部位的疱疹、斑丘疹,其病原体主要是EV71和柯萨奇A组16型COX16〔1〕。最早在1957年由新西兰Sedno首次报告,1958年加拿大Robinson从患儿粪便和咽拭子标本中分离出柯萨奇病毒,1959年在英国伯明翰流行时Alsop氏首先提出HFMD命名。1969年美国首次从加利福尼亚患有中枢神经系统疾病的婴儿粪便标本中分离出肠道病毒71型(EV71)。自此,EV71已在世界范围内引起多次暴发流行,近年在亚太地区也呈上升趋势。柯萨奇病毒A16型与肠道病     

2013-2014年泉州地区手足口病病原学监测结果分析

摘要:目的　了解2013-2014年福建泉州地区手足口病（HFMD）的病原学特征,为手足口病防治提供科学依据。方法　采集2013-2014年泉州市妇幼保健院·儿童医院收治的3839例手足口病患儿空腹静脉血标本,采用捕获法酶联免疫吸附技术检测肠道病毒71型（EV71-IgM）和柯萨奇病毒A组16型（CA16-IgM）。结果　3839例被检标本共检测出阳性标本2176例,阳性率为56.68%;其中32.87%（1262/3839）为EV71阳性,23.81%（914/3839）为CA16阳性,两者间阳性率差异有统计学意义（P＜0.05）。5岁以下HFMD患者3679例,占95.83%,其中1～2岁组患儿最多,占68.56%。3～4岁组患儿EV71阳性率（48.28%）和CA16阳性率（35.09%）最高。不同年龄组,EV71阳性率和CA16阳性率均存在统计学差异（P＜0.05）。手足口病发病随季节而发生变化,不同月份EV71和CA16阳性率均存在统计学差异（P＜0.05）。发病以4-7月份为主,占56.37%,2014年9-10月份发病率出现一定程度反弹。结论　2013-2014年泉州地区手足口病肠道病毒EV71型的阳性检出率高于CA16型,4-7月份为发病高峰期,5岁以下儿童为高发人群,其中1～2岁组手足口患儿居多,而3～4岁组病毒阳性率最高,需引起重视。     

A novel inactivated enterovirus 71 vaccine can elicit cross-protective immunity against coxsackievirus A16 in mice

Hand, foot, and mouth disease (HFMD) is a highly contagious disease that mainly affects infants and children. Enterovirus 71 (EV71) and coxsackievirus A16 (CA16) are the major pathogens of HFMD. Two EV71 vaccines were recently licensed in China and the administration of the EV71 vaccines is believed to significantly reduce the number of HFMD-related severe or fatal cases. However, a monovalent EV71 vaccine cannot cross-protect against CA16 infection, this may result in that it cannot effectively control the overall HFMD epidemic. In this study, a chimeric EV71, whose VP1/210–225 epitope was replaced by that of CA16, was constructed using a reverse genetics technique to produce a candidate EV71/CA16 bivalent vaccine strain. The chimeric EV71 was infectious and showed similar growth characteristics as its parental strain. The replacement of the VP1/210–225 epitope did not significantly affect the antigenicity and immunogenicity of EV71. More importantly, the chimeric EV71 could induce protective immunity against both EV71 and CA16, and protect neonatal mice against either EV71 or CA16 lethal infections, the chimeric EV71 constructed in this study was shown to be a feasible and promising candidate bivalent vaccine against both EV71 and CA16. The construction of a chimeric enterovirus also provides an alternative platform for broad-spectrum HFMD vaccines development.     

河北省2009年手足口病病原学研究

目的了解2009年引起河北省手足口病的病原谱及分布特征,为制定预防控制措施提供实验依据。方法通过流行病学调查,采集手足口病临床诊断病例的粪便、肛拭子、咽拭子、疱疹液和脑脊液标本,针对肠道病毒(EV)5′端非编码区保守基因序列以及EVVP1至2A区基因序列设计EV通用引物和型特异性引物,通过RT-PCR扩增检测HFMD临床诊断病例的标本,并对病原进行分型。结果采用RT-PCR法共检测HFMD临床诊断病例1990份标本,1296份标本EV阳性,阳性率为65.13%,其中752份标本EV71阳性,452份CA16阳性,1份EV71+CA16阳性,91份其他EV阳性。随机抽取55份其他EV阳性标本经RT-PCR和测序进一步分型,除24份未定型外,共鉴定出31份,其中CA10占18.18%(10/55),CA6占14.55%(8/55),ECO30占9.09%(5/55),ECO6、ECO9各占3.64%(2/55),CA5、CA14、CB5、ECO14各占1.82%(1/55),未定型占43.64%(24/55)。结论 2009年引起河北省手足口病流行的病原体主要为EV71和CA16,同时还有部分CA5、CA6、CA10、CA14、CB5、ECO6、ECO9、ECO14、ECO30等其他型别,引起河北省重症病例的EV主要型别为EV71。     

Hepatitis B virus core particles containing multiple epitopes confer protection against enterovirus 71 and coxsackievirus A16 infection in mice

Human enterovirus 71 (EV71) and coxsackievirus A16 (CA16) are the two major causative agents of hand-foot-and-mouth disease (HFMD). To investigate novel combined vaccines to prevent EV71 and CA16 infection, we constructed chimeric virus-like particles (tHBc/SPA or tHBc/SP VLPs) displaying conserved epitopes of EV71 (aa 208–222 of VP1 and aa 248–263 of VP2) and CA16 (aa271-285 of VP1) using a truncated hepatitis B virus core carrier (tHBc). Immunization with the chimeric VLPs induced epitope- or virus-specific IgG and neutralization antibodies against EV71 and CA16 in the mice. Compared with inactivated EV71, the chimeric VLPs induced significantly increased Th1 cytokine (IFN-γ, IL-2) production and decreased Th2 cytokine (IL-4, IL-10) responses. Neonatal mice born to dams immunized with the recombinant particles were completely protected from lethal EV71 and partially protected from CA16 infection. Co-expression of the conserved human MHC class I CD4+ T cell epitope (aa248-263 of VP2) did not improve the antiviral immunity of the chimeric VLP vaccine in mice. Our results demonstrate that experimental combination vaccines comprised of EV71 and CA16 epitopes induce both humoral and cellular immune responses and therefore support further preclinical and clinical development of a bivalent VLP vaccine targeting both CA16 and EV71.     

2010年合肥儿童手足口病病例肠道病毒血清基因型分析

目的:对合肥地区2010年5~7月儿童手足口病流行期间的手足口病患儿进行病原学调查。方法:采集手足口病患儿的疱疹液、粪便、咽拭子进行病毒分离;分离的病毒接种非洲绿猴肾细胞(Vero)后待细胞病变(CPE)达到++++以上后提取RNA。分别用EV71和CA16的VP1基因的特异性引物进行RT-PCR鉴定,并根据血清型不同比较其临床特征。结果:77例患儿采集的标本中65例分离到病毒;经RT-PCR检测发现其中47例CA16阳性,18例为EV71,比例为2.6∶1。CA16感染和EV71感染在患者的年龄大小、症状轻重等方面无显著统计学差异。结论:手足口病患儿检测到的主要病原是CA16和EV71,两者感染对临床的影响无显著差异。     

A combination vaccine comprising of inactivated enterovirus 71 and coxsackievirus A16 elicits balanced protective immunity against both viruses

Enterovirus 71 (EV71) and coxsackievirus A16 (CA16) are the two major causative agents of hand, foot and mouth disease (HFMD), which is an infectious disease frequently occurring in children. A bivalent vaccine against both EV71 and CA16 is highly desirable. In the present study, we compare monovalent inactivated EV71, monovalent inactivated CA16, and a combination vaccine candidate comprising of both inactivated EV71 and CA16, for their immunogenicity and in vivo protective efficacy. The two monovalent vaccines were found to elicit serum antibodies that potently neutralized the homologous virus but had no or weak neutralization activity against the heterologous one; in contrast, the bivalent vaccine immunized sera efficiently neutralized both EV71 and CA16. More importantly, passive immunization with the bivalent vaccine protected mice against either EV71 or CA16 lethal infections, whereas the monovalent vaccines only prevented the homologous but not the heterologous challenges. Together, our results demonstrate that the experimental bivalent vaccine comprising of inactivated EV71 and CA16 induces a balanced protective immunity against both EV71 and CA16, and thus provide proof-of-concept for further development of multivalent vaccines for broad protection against HFMD.     

Hexon-modified recombinant E1-deleted adenoviral vectors as bivalent vaccine carriers for Coxsackievirus A16 and Enterovirus 71

Hand, foot and mouth disease (HFMD) is a major public health concern in Asia; more efficient vaccines against HFMD are urgently required. Adenoviral (Ad) capsids have been used widely for the presentation of foreign antigens to induce specific immune responses in the host. Here, we describe a novel bivalent vaccine for HFMD based on the hexon-modified, E1-deleted chimpanzee adenovirus serotype 68 (AdC68). The novel vaccine candidate was generated by incorporating the neutralising epitope of Coxsackievirus A16 (CA16), PEP71, into hypervariable region 1 (HVR1), and a shortened neutralising epitope of Enterovirus 71 (EV71), sSP70, into HVR2 of the AdC68 hexon. In order to enhance the immunogenicity of EV71, VP1 of EV71 was cloned into the E1-region of the AdC68 vectors. The results demonstrated that these two epitopes were well presented on the virion surface and had high affinity towards specific antibodies, and VP1 of EV71 was also significantly expressed. In pre-clinical mouse models, the hexon-modified AdC68 elicited neutralising antibodies against both CA16 and EV71, which conferred protection to suckling mice against a lethal challenge of CA16 and EV71. In summary, this study demonstrates that the hexon-modified AdC68 may represent a promising bivalent vaccine carrier against EV71 and CA16 and an epitope-display platform for other pathogens.     

Chimeric enterovirus 71 virus-like particle displaying conserved coxsackievirus A16 epitopes elicits potent immune responses and protects mice against lethal EV71 and CA16 infection

Hand-foot-and-mouth disease (HFMD) is an infectious disease of infants and young children frequently caused by the enterovirus A species, mainly enterovirus 71 (EV71) and coxsackievirus A16 (CA16). In this study, we prepared the EV71 virus-like particle (EV71-VLP) and its chimeras using recombinant baculovirus (Bac-P1-3CD) co-expressing EV71 P1 (under polyhedrin promoter) and 3CD (under CMV-IE promoter) proteins in Sf9 cells. EV71-VLP chimera ChiEV71(1E)-VLP or ChiEV71(4E)-VLP displayed single CA16 PEP71 epitope in VP1 or four conserved CA16 neutralizing epitopes (PEP71 in VP1, aa136-150 in VP2, aa176-190 in VP3 and aa48-62 in VP4) by substitution of the corresponding regions of EV71 structure proteins, respectively. In mice, EV71-VLP and its chimeras elicited similar EV71-specific IgG and neutralizing antibody (NAb) titers compared to inactivated EV71. Expectedly, vaccination of ChiEV71(1E)-VLP or ChiEV71(4E)-VLP resulted in significantly increased CA16-specific IgG and NAb production and improved cross-protection against CA16 infection compared to EV71-VLP. Interestingly, the VLPs induced potent cellular immune responses and significantly decreased Th2 type (IL-4 and IL-10) cytokines secretion in the splenocytes of immunized mice compared to inactivated EV71 or inactivated CA16. Neonatal mice born to dams immunized with the chimeric VLPs or neonatal mice passively transferred with sera of immunized mice were completely protected from lethal EV71 challenge and partially protected from lethal CA16 infection. Our study provides a novel bivalent or multivalent vaccine strategy to prevent EV71 and related-enterovirus infections.     

北京及其周边地区儿童手足口病病原学监测及流行病学分析(2017-2018年)

目的了解北京及其周边地区儿童手足口病(HFMD)的流行特点及病原学特征。方法采集北京及其周边地区2017—2018年儿童HFMD病例咽拭子样本,使用肠道病毒(EV)通用、EV71及柯萨奇病毒A16型(CA16)三通道Real-time PCR的方法进行肠道病毒核酸检测,对非EV71非CA16的EV阳性样本采用RT-PCR法扩增VP1基因片段,通过BLAST比对进一步进行型别鉴定。结果2017—2018年共纳入HFMD患者939例,包括重症病例2例;男女性别比为1.54∶1;3岁以下儿童占46.96%,5岁以下儿童占75.19%。EV总阳性检出率为89.24%(838/939),其中2017年、2018年分别为87.44%(181/207)和89.75%(657/732);2017年主要为CA6(47.51%)、EV71(16.02%)、CA16(7.73%)和未分型EV(27.62%);2018年包括CA6(46.88%)、CA16(14.31%)和未分型EV(36.99%)。结论2017—2018年,引起北京及其周边地区儿童HFMD的主要病原体是CA16,EV71的检出率显著下降,因此需要加强HFMD病原体中CA6等非EV71非CA16肠道病毒的监测工作。