2011年厦门市某医院十例无菌性脑炎暴发病例病原学分析

目的 对2011年5月11-17日厦门某医院收治的10例无菌性脑炎患者进行病原学鉴定.方法 采集10例患者的咽拭子、肛拭子及部分患者的脑脊液,进行总肠道病毒核酸检测.总肠道病毒核酸阳性的样本,分别用肠道病毒A、B、C基因型各自的VP1通用引物扩增,VP1区扩增阳性的标本测序后进行序列分析.同时,用Vero细胞进行病毒分离培养,对培养成功的病毒与原始临床标本进行VP1序列同源性分析,从而对引发本次脑炎的病原进行鉴定.结果 10例脑炎患者中,有7例患者的临床标本总肠道病毒核酸检测阳性.这7例患者的咽拭子和(或)肛拭子标本肠道病毒B基因型VP1通用引物扩增阳性,测序后分析表明均为肠道病毒B基因型埃可病毒30(Echo 30),且与浙江2004年流行毒株的同源性达95.3%～97.1%.其中编号为XM2、XM3、XM4、XM8的咽拭子和XM3、XM6的肛拭子相互间序列同源性为99.4%～100.0%,但与XM1的咽拭子序列存在差异,同源性为92.8%～93.4%.此外,XM1、XM2、XM3、XM4、XM8咽拭子标本Vero细胞分离培养病毒成功,其VP1区部分序列与相应的原始标本同源性大于99.9%.结论 导致这次无菌性脑炎的病原体为肠道病毒B基因型Echo 30,并提示可能存在多种不同遗传背景的Echo 30在流行.

Abstract：

Objective To identify the etiology of an aseptic encephalitis outbreak (ten cases) in a hospital of Xiamen city from 11 to 17 May, 2011.Methods A total of ten patients′ throat swabs, anal swabs and cerebrospinal fluid were collected and detected by RT-PCR for pan-enterovirus. The samples containing detectable pan-enterovirus were tested by PCR with genotype-specific general primers located in VP1 region of enterovirus genotype A, B and C (HEV-A, B and C). The PCR products of VP1 segment were purified and sequenced, and phylogenetic analysis was performed. Meanwhile, the pathogens in those samples were isolated in Vero cell culture. Homologous analysis of VP1 sequences were carried out for the cultured virus samples and the original clinical samples to identify the outbreak etiology.Results Among the ten cases, seven cases were positive for pan-enterovirus nucleic acid. When tested by genotype-specific PCR, the throat and anal swab samples from those 7 patients were positive with HEV-B VP1 primers. Meanwhile, the HEV-B VP1 segments were sequenced and phylogenetic analyzed, which indicated the seven cases were all infected by enterovirus Echo 30. The sequences from those samples had homology of 95.3%-97.1% with the epidemic strains in Zhejiang, 2004. Out of the seven cases, the sequences of XM2,XM3,XM4,XM8 throat swab samples and XM3,XM6 throat samples showed 99.4%-100.0% homology which were different from the sequence of XM1, and the homology was 92.8%-93.4%. Furthermore, the viruses were isolated using Vero cells from XM1,XM2,XM3,XM4 and XM8 throat swab samples,and the VP1 sequence showed more than 99.9% homology with the original specimens.Conclusion The local outbreak of aseptic encephalitis was caused by Echo 30 of enterovirus genotype B, and the epidemic strains may have different genetic background.     

云南省2007--2009年流行性腮腺炎病毒SH和HN基因的遗传特征分析

目的 分析2007-2009年云南省腮腺炎病毒(MuV)分离株的SH和HN遗传特征.方法 采用反转录-聚合酶链反应从Vero细胞分离的病毒株基因组中扩增出SH基因和HN基因,测序后用Mega4.1软件分析其遗传特征.结果 云南省分离14株MuV的SH基因其核苷酸和氨基酸同源性为98.3%～100.0%和96.5%～100.0%,与其他省份比较其同源性为92.6%～99.4%和87.7%～100.0%,其中Wsh1和Wsh2与其他F基因型差异较大;与疫苗株同源性为84.5%～85.1%和77.2%;与其他基因型同源性为83.4%～90.9%和70.1%～86.0%.6株MuV分离株的HN基因与核苷酸和氨基酸同源性分别为99.3%～99.5%和99.1%～99.7%;与中国分离株SP株的核苷酸和氨基酸同源性均为99.8%;与其他基因型同源性为94.7%～96.8%和95.5%～99.1%;与疫苗株的同源性为92.4%～93.2%和95.5%～96.4%.结论 2007-2009年云南省流行的MuV均为F基因型;其HN基因比SH基因保守.

Abstract：

Objective To analyze genetic characterization of the small hydrophobic and hemagglutinin-neuraminidase genes of mumps virus(MuV)isolated in Yunnan province,China from 2007 to 2009.Methods Fourteen MuV strains were isolated in Yunnan,China from 2007 to 2009.Using RT-PCR,the SH gene fragments contained 316 nucleotides in all strains and HN gene of six strains were sequenced.The sequences were aligned with other mumps virus sequences downloaded from GenBank using Mega 4.1 software.Results Fourteen isolated strains were closely related to other reference strains of F genotypes.In SH gene,the homology of nucleotide and amino acid among the fourteen isolated strains were 98.3%-100.0%and 96.5%-100.0%,respectively,and 92.6%%-99.4%and 87.7%-100.0% of homology when compared with that of strains isolated from other provinces in China,respectively.Wsh1 and Wsh2 strains had less homology when compared to other strains of F genotypes.The fourteen strains had homology of 84.5%-85.1%and 77.2%compared to vaccine strains on nucleotide and amino acid,respectively,and had homology of 83.4%-90.9% and 70.1%-86.0% compared to that of other genotypes.In HN gene,the homology of nucleotide and amino acid among the six isolated strains were 99.3%-99.5% and 99.1%-99.7%,respectively,and also 99.8% and 99.8% of homology respectively when compared to the SP strain in China.All the six strains had homology of 92.4%-93.2% and 95.5%-96.4% when compared to the vaecine strains on nucleotide and amino acid,respectively,and had homology of 94.7%-96.8% and 95.5%-99.1%compared to other genotypes.Conclusion Fourteen strains isolated in Yunnan from 2007 to 2009belonged to F genotype of MuV while the HN gene seemed more conservative than SH gene.     

浙江省临海市2004年病毒性脑膜炎暴发疫情的病原学及其分子特征研究

目的 为确证2004年浙江省临海市病毒性脑膜炎暴发疫情的病因,分析病原的遗传变异及进化关系.方法 采集患者脑脊液样本60份,采用RD和Hep-2细胞同时分离病毒,中和试验法鉴定病毒型别;对分离株VP1和VP4/VP2基因测序,进行同源性与进化分析.结果 从60份脑脊液样本中分离到埃柯病毒30型(E30)19株,分离率为31.7％;对4株E30分离株VP1区核苷酸序列测定,其长度均为876个核苷酸(nt),编码292个氨基酸(aa).临海分离株与E30原型株Bastianni在VP1区的nt和aa同源性分别为82.4％～84.1％和93.5％ ～ 94.2％;4株临海E30株之间nt和aa的同源性分别为87.1％～99.9％和97.9％ ～ 100.0％.临海分离株分成两类,同类病毒株间的差异很小,而两类病毒株间的差异很大,nt和aa的最大差异率分别为12.9％和2.1％.与临海E30株同源性最高的为2002-2003年浙江E30株.在VP1基因进化树上,临海E30株分别位于G和H基因亚型分支上,其中临海G基因株与2003年浙江、江苏和山东E30株位于同一进化分支,临海H基因株与2002年浙江诸暨株位于同一进化分支.VP4/VP2区进化分析结果与VP1区相似.结论 2004年临海市病毒性脑膜炎暴发疫情由E30G和H不同基因亚型的E30流行株引起;临海E30株与2002-2003年浙江、江苏和山东E30株具有密切的亲缘关系;H基因亚型株推测为新的E30变异株,首先分离于2002年浙江省.     

2008年和2010年肠道病毒7l型广州分离株全基因组序列分析

目的 研究2008年和2010年广州地区手足口病患者中肠道病毒71型(EV71)病毒全基因组序列的基因型与变异特点.方法 参照GenBank上EV71深圳株SHZH03(AY465356)基因组设计分段扩增引物,进行RT-PCR分段扩增EV71病毒基因组,PCR产物直接进行序列测定,用Clustal W/X、DNASTAR、MEGA4.1等软件分析基因组序列.结果 克隆9株广州株EV71,病毒全基因组全长序列均为7405 bp,提交到GenBank上的序列号为HQ456305、HQ456306、HQ456307、HQ456308、HQ456309、HQ456310、HQ456311、HQ456312、HQ456313.将9株广州株EV71与EV71的A、B3、B4、B5、C1、C2、C3、C4型及阜阳株全基因组核苷酸序列进行Clustal W比较,发现与C4a型阜阳株等同源性为98%～99%,与C4b同源性为91%～93%,与C1～C3同源性为82%～83%,与B3～B5同源性为81%～83%,与A型同源性为80%.将9株广州分离株EV71的VP1基因与EV71病毒A、B、C型进行Clustal W比较,同样是与C4a为98%～99%,与C4b同源性为92%～94%,与C1～C3的同源性为88%～89%,与B1～B5型同源性为83%～84%,与A型同源性为81%～82%.将9株广州株与EV71的A、B、C型VP1基因氨基酸序列进行Clustal W比对,发现VP1基因22位点的谷氨酰胺转变为组氨酸(Q→H),聚合蛋白中213位点(S→T)和1764位点(V→(Ⅰ)也发生了氨基酸的点突变,P的213位点属于VP2基因,1764位点属于3D基因.结论 2008年和2010年分离的9株广州株EV71属于C4a型,与阜阳株同源性为98%～99%,VP1基因22位点发生了点突变,聚合蛋白中213位点和1764位点也发生了氨基酸的点突变.

Abstract：

Objective To study the genomic genotypes and variation of human enterovirus 71(EV71)infected infants in Guangzhou city,in 2008 and 2010.Methods Primers were designed on the basis of the genomic sequence of EV71 SHZH03 strain(AY465356)in the GenBank,and EV71genome amplified by RT-PCR.PCR-products were directly sequenced and the genomic nucleotide sequences were analyzed with the programs of Clustal W/X,DNASTAR and MEGA 4.1.Results 9strains of EV71 genome appeared to be 7405 bp in length.The genomic sequences of EV71Guangzhou strains were compared with those of EV71 in GenBank,which revealed that the homology with EV71 genotype C4a Fuyang strains ranged between 98%-99%.Homology with genotype C4b were 92%-94%,with genotypes C1,C2,C3 as 82%-83%,with genotypes B3,B4,B5 as 81%-83%and the homology with genotype A was 80%.When compared the VP1 genes of EV71 Guangzhou strains with genotypes A,B,C virus,we revealed that the highest homology was also with genotype C4a.When compared the VP1 amino acid sequences of EV71 Guangzhou strains with genotype A,B,C virus by Clustal W program,the results revealed that the amino acid residue Q at position 22 in VP1gene was transformed to H,while 213(S→T)and 1764(V→(Ⅰ))mutations in polyprotein were discovered.Conclusion Data from the sequences and phylogenetics analysis on those Guangzhou strains in 2008 and 2010 revealed that those isolates belong to genotype C4a,with the homology with Fuyang strains as 98%-99%.Mutation of amino acid residue H at position 22 in VP1 gene was discovered and the neutralizing antibody of EV71 might have been conversed by this residue.213(S→T)and 1764(V→Ⅰ)mutations in polyprotein were also discovered.     

浙江省病毒性脑炎病原柯萨奇B1病毒分子特征研究

目的:为了查明引起2008-2009年浙江省疑似病毒性脑炎的病原,分析病原的分子特征。方法:采用Hep-2和RD细胞从疑似患者脑脊液样本中分离肠道病毒,对分离株扩增VP1基因,测序并进行同源性与进化分析。结果:从患者11份脑脊液样本中分离到4株柯萨奇B1病毒(CVB1);测得VP1基因全长为834核苷酸(nt),无插入和缺失,推导编码278个氨基酸(aa)。4株浙江CVB1分离株(ZJ/CVB1)之间在VP1区nt和aa的同源性分别为96.3%~99.9%和100%。浙江CVB1分离株BLAST比对nt同源性最高的毒株为2006年山东株(SD/168/06),同源性为98.7%;与CVB1原型株(Conn-5/86)的nt同源性为78.9%。VP1区氨基酸进化树显示浙江CVB1分离株在CVB1 Clade3进化支上,与浙江株亲缘关系最近的毒株为山东CVB1株,均在同一进化支上。结论:从病毒分离结果证实浙江省疑似病毒性脑炎是由肠道病毒CVB1引起,2008-2009年在浙江循环的CVB1病毒之间没有明显差异,浙江CVB1流行株与山东株亲缘关系最近。     

华东和华南地区商品猪戊型肝炎病毒携带率和基因分型分析

Objective To determine the prevalence and genotype of hepatitis E virus (HEV)among commercial swine population in Eastern and Southern China. Methods Six hundred specimens of swine bile collected from 5 slaughterhouses in Eastern and Southern China from 2007 to 2009 were tested for HEV RNA using nested RT-PCR. PCR products were sequenced for phylogenetic analysis. Results Forty-seven out of the 600 samples (7.83%) were positive for HEV RNA. Based on the 150 nt fragment within HEV ORF2, data from phylogenetic analysis revealed that all the 47 HEV isolates were identified to be genotype Ⅳ, sharing 75.0%-83.4%, 75.0%-84.6%, 71.9%-80.7% and 88.1%-91.5% nucleotide identities with prototype Ⅰ,Ⅱ,Ⅲ and Ⅳ HEV strains respectively while majority of the isolates clustered within their respective isolation sites. Conclusion HEV was widespread in commercial swine population in Eastern and Southern China that raised a serious concern about the safety regarding the consumption of pork products.     

华东和华南地区商品猪戊型肝炎病毒携带率和基因分型分析

Objective To determine the prevalence and genotype of hepatitis E virus (HEV)among commercial swine population in Eastern and Southern China. Methods Six hundred specimens of swine bile collected from 5 slaughterhouses in Eastern and Southern China from 2007 to 2009 were tested for HEV RNA using nested RT-PCR. PCR products were sequenced for phylogenetic analysis. Results Forty-seven out of the 600 samples (7.83%) were positive for HEV RNA. Based on the 150 nt fragment within HEV ORF2, data from phylogenetic analysis revealed that all the 47 HEV isolates were identified to be genotype Ⅳ, sharing 75.0%-83.4%, 75.0%-84.6%, 71.9%-80.7% and 88.1%-91.5% nucleotide identities with prototype Ⅰ,Ⅱ,Ⅲ and Ⅳ HEV strains respectively while majority of the isolates clustered within their respective isolation sites. Conclusion HEV was widespread in commercial swine population in Eastern and Southern China that raised a serious concern about the safety regarding the consumption of pork products.     

1998-2009年浙江省H3N2亚型流行性感冒病毒全序列变异的研究

目的 比较1998-2009年浙江省H3N2亚型流行性感冒(简称流感)流行株HA基因进化与全基因进化状况的一致性,并探讨全基因组序列上可能存在的潜在抗原区域.方法 选择浙江省CDC流感实验室于1998-2009年流感疫情中分离保存的H3N2亚型流感流行代表株19株,采用RT-PCR法进行全基因组序列扩增,并将这些毒株与10株同期H3N2亚型流感疫苗株进行全序列及HA1基因的系统进化比较;通过氨基酸替换比较、熵值计算及正向选择位点的筛选,确定各基因上的氨基酸易变位点.结果 H3N2亚型流感病毒的全序列长度为4466个氨基酸,其中有137个氨基酸位点发生稳定变异.在HA基因上第144和158位氨基酸分别经历了4次和3次替换,NA基因的第93、143、307、370、372位氨基酸和NP基因的第450位氨基酸经历了2次替换,且HA基因和NA基因上分别有29%(12/41)和77%(24/31)的变异位点位于已知抗原决定簇之外的区域.氨基酸位点熵值分析显示,HA基因非抗原决定簇的3、225、361位,NA基因非抗原决定簇的93、143、147、150、372位,PB1基因的113、576、586位,PA基因的101、256、382、421、437位,NP基因的377、450位,M1基因的218位及M2基因的31位氨基酸均为易变位点.结论 浙江省1998-2009年H3N2亚型流感病毒在HA和NA已知抗原决定簇之外的区域与部分内部基因上,可能存在着一些尚未被发现或者新形成的抗原位点.与HA基因的系统进化比较,全序列能更为全面地反映出流感流行株之间的亲缘关系与进化规律.

Abstract：

Objective To analyze the consistency of evolution condition between HA gene and the whole genome of influenza virus subtype A/H3N2 strains isolated in Zhejiang province from 1998 to 2009,and to study the potential antigenic region on the whole genome.Methods The sequences of whole genome of 19 Zhejiang influenza virus isolates circulated from 1998 to 2009,which conserved by influenza laboratory of Zhejiang Provincial Centre for Disease Prevention and Control,were amplified using RT-PCR assays.The obtained sequences were used to conduct phylogenetic analysis with 10 contemporaneous vaccine strains.Three methods,including comparison of the amino acid substitutions,calculation of the entropy value and the filtering of positive selection sites,were used to confirm the mutable sites on each gene.Results The whole genome of influenza virus subtype A/H3N2 was 4466 amino acids in length,with 137 stable mutations.The 144,158 aa of HA gene mutate four and three times respectively; 93,143,307,370,372 aa of NA gene and 450 aa of NP gene mutate twice,and there were 29%(12/41) and 77%(24/31) mutations of HA and NA genes occurred on the non-epitope regions respectively.Analysis of the entropy value suggest that many amino acid sites on the non-epitope regions were prone to mutation,including 3,225,361 aa of HA gene; 93,143,147,150,372 aa of NA gene; 113,576,586 aa of PB1 gene; 101,256,382,421,437 aa of PA; 377,450 aa of NP gene; 218 aa of M1 gene and 31 aa of M2 gene.Conclusion Based on the whole genome of influenza virus subtype A/H3N2 strains isolated in Zhejiang province in 1998 to 2009,there may be several unknown or new antigen sites existing on the non-epitope regions of HA and NA genes and parts of internal genes.The phylogenetic tree constructed based on the complete sequence was more comprehensive than on the HA gene to reflect the genetic relationship and law of evolution among the influenza virus strains.     

开封地区病毒性脑炎病原分离株Echo30的分子流行病学分析

目的研究开封地区2008年病毒性脑炎病原分离株Echo30VP1区基因特征及其分子流行病学特点。方法对开封地区2008年病毒性脑炎监测病例脑脊液标本中分离到的8株Echo30病毒进行VP1区全基因序列测定,与GeneBank上发表的ECHO30型VP1区进行同源性比较,通过构建进化树对其进行遗传进化分析。结果所测8株Echo30VP1区基因全长均为876bp,翻译的氨基酸全长292aa。8株病毒VP1区核苷酸同源性为92.8%～99.7%,氨基酸同源性为93.0%～99.7%;与GenBank相关毒株基因组序列比对显示开封地区2008年分离株均与江苏省2003年分离株同源性较高;进化分析表明属于第7组。结论 2008年开封地区Echo30分离株与2003年江苏省分离株亲缘关系最近,可能具有相同来源。     

Identification of a novel canine norovirus

By screening a collection of fecal samples from young dogs from different European countries, noroviruses (NoVs) were found in 13/294 (4.4%) animals with signs of enteritis whilst they were not detected in healthy dogs (0/42). An informative portion of the genome (3.4 kb at the 3′ end) was generated for four NoV strains. In the capsid protein VP1 region, strains 63.15/2015/ITA and FD53/2007/ITA were genetically related to the canine GVI.2 strain C33/Viseu/2007/PRT (97.4–98.6% nt and 90.3–98.6% aa). Strain FD210/2007/ITA displayed the highest identity to the GVI.1 canine strain Bari/91/2007/ITA (88.0% nt and 95.0% aa). Strain 5010/2009/ITA displayed only 66.6–67.6% nt and 75.5–81.6% aa identities to the GVI.1 canine strains FD210/2007/ITA and Bari/91/2007/ITA and the GVI feline strain M49-1/2012/JPN. Identity to the other canine/feline NoVs strains in the VP1 was lower than 67.6% nt and 62.7% aa. Based on the full-length VP1 amino acid sequence and the criteria proposed for distinction of NoV genotypes, the canine NoV 5010/2009/ITA could represent the prototype of a third GVI genotype, thus providing further evidence for the genetic heterogeneity of NoVs in carnivores.     

2009—2017年柯萨奇病毒A组4型贵州分离株的基因特征分析

目的 分析贵州省柯萨奇病毒A组4型（coxsackievirus A4，CVA4）分离株的基因特征。方法 对2009—2017年贵州省急性弛缓性麻痹（acute flaccid paralysis, AFP）病例中分离到的19株CVA4，采用反转录-聚合酶链反应（RT - PCR）方法进行VP1区扩增并对扩增产物测序，通过MEGA 7.0软件采用邻位相接法（neighbor joining method）构建遗传进化树进行基因特征分析，可靠性通过1 000 Bootstrap值评估。结果 19株CVA4贵州分离株之间的的核苷酸同源性为92.0%～100.0%，氨基酸同源性为96.1%～100.0%，与原型株high point株之间的核苷酸同源性为83.3%～85.7%，氨基酸同源性为95.1%～98.4%； 19株CVA4均为C2基因亚型，主要聚集在Cluster1和Cluster 2分支，其中Cluster1是2009年后贵州地区持续传播的优势群体。结论 2009—2017年贵州省AFP病例中CVA4贵州分离株为C2基因亚型。     

济南市轻重症手足口病患儿肠道病毒71型结构蛋白基因特征分析

目的 比较分析济南市轻重症手足口病(hand, foot and mouth disease, HFMD)病原肠道病毒71型(enterovirus 71, EV71)结构蛋白基因及氨基酸变异情况。方法 对2017年济南市轻重症手足口病患儿EV71细胞分离株结构蛋白基因序列进行RT-PCR扩增测序,应用EditSeq和MegAlign等软件对结构蛋白基因进行序列拼接及核苷酸、氨基酸同源性比对,应用Mega 5.2软件构建VP1基因系统进化树。结果 轻重症患儿EV71分离株结构蛋白基因核苷酸和氨基酸同源性分别为93.0%～98.8%和99.0%～99.8%,均属于C4a基因亚型。经VP1基因同源性分析,济南市EV71分离株与2017年江苏株（GenBank ID: MG520666,同源性99.3%）、2015年河南株（KU744452,同源性99.2%）、2015年北京株（KU376386,同源性99.2%）及2016年山东潍坊株（MG588036,同源性99.0%）亲缘关系较近。死亡患儿EV71分离株VP1蛋白出现E145Q突变,重症EV71分离株分别出现VP1蛋白A19V、T101I,VP3蛋白M150I突变。结论　济南市轻重症HFMD患儿EV71分离株均属于C4a亚型,与江苏、河南、山东及北京等地的EV71株亲缘关系较近,其VP1蛋白145位氨基酸残基可能是影响病毒毒力的重要位点。     

A novel feline norovirus in diarrheic cats

By screening a collection of fecal samples from young cats housed in three different shelters in South Italy, noroviruses (NoVs) were found in 3/48 (6.2%) specimens of animals with enteritis signs while they were not detected in samples collected from healthy cats (0/57). Upon sequence analysis of the short RNA-dependent RNA polymerase (RdRp) region, the three strains displayed the highest nucleotide (nt) and amino acid (aa) identities to the prototype GIV.2 strain lion/Pistoia/387/06/ITA (91.0–93.0% nt and 97.0–98.0% aa). The sequence of ~ 3.4-kb portion at the 3′ end of the genome of a NoV strain, TE/77-13/ITA, was determined. In the full-length ORF2, encoding the VP1 capsid protein, the virus was genetically closest to the canine GVI.2 NoV strains C33/Viseu/2007/PRT and FD53/2007/ITA (81.0–84.0% nt and 93.0–94.0% aa identities), suggesting a recombination nature, with the cross-over site being mapped to the ORF1-ORF2 junction. Based on the full-length VP1 amino acid sequence, we classified the novel feline NoV, together with the canine strains Viseu and FD53, as a genotype 2, within the genogroup GVI. These findings indicate that, as observed for GIV NoV, GVI strains may infect both the canine and feline host. Unrestricted circulation of NoV strains in small carnivores may provide the basis for quick genetic diversification of these viruses by recombination. Interspecies circulation of NoVs in pets must also be considered when facing outbreaks of enteric diseases in these animals.     

2009—2019年上海市闵行区手足口病病原谱及EV-A71和CV-A16病毒分离株VP1区基因特征分析

目的 了解2009—2019年上海市闵行区手足口病的病原谱,分析肠道病毒71型(enterovirus 71,EV-A71)和柯萨奇病毒A组16型(coxsackievirus A16,CV-A16)VP1区基因特征,为手足口病的综合防治提供科学依据。方法 对2009—2019年闵行区手足口病监测点送检的标本应用实时荧光定量PCR进行病原学检测,分析病原学特征。EV-A71和CV-A16病毒分离株进行VP1区核苷酸序列测定,分析其同源性并构建系统进化树。结果 2009—2019年共收集到5 364例手足口病病例标本,病原学检测阳性检出率为86.74%(4 653/5 364),其中EV-A71检出率为40.12%(2 152/5 364)、CV-A16为16.61%(891/5 364)、CV-A6为20.86%(1 119/5 364)、CV-A10为1.49%(80/5 364)。2009—2014年主要呈EV-A71和CV-A16共同流行趋势,2015—2019年主要以CV-A6和CV-A16共同流行为主。EV-A71分离株均为C4a型,VP1区核苷酸序列同源性为92.1%～99...     

北京地区2006-2008年肠道病毒71型VP1区基因特征分析

目的 了解2006-2008年北京地区分离的不同来源肠道病毒71型(EV71)VP1区基因特征.方法 从手足口病例、急性弛缓性麻痹病例和健康儿童的粪便、咽拭子和疱疹液标本中分离EV71病毒株,选取其中9株EV71分离株,采用RT-PCR扩增VPl区全长基因片段并进行序列测定、系统发生树构建、核苷酸同源性及组内和组间进化距离分析.结果 9株EV71在系统发生树上与C基因型中的C4亚型代表株属同一分支;在核苷酸同源性分析中,9株分离株与C4亚型代表株的同源性达到92.1%～93.9%,明显高于与C1、C2、C3亚型代表株的同源性(分别为88.8%～89.5%、89.4%～90.0%和88.4%～89.3%);三种不同来源的分离株病毒之间具有较高的同源性(95.9%～100.0%),但与1998年分离的C4亚型代表株的同源性却较低(分别为93.3%～93.9%和92.1%～92.9%).在进化距离的分析上,C4亚型内各组间距离较大,尤其是亚型代表株与分离株之间距离最大(D=0.052～0.071).结论 2006-2008年北京地区流行的EV71仍然是C4亚型,在同一时间不同地区、不同来源、不同疾病状态卞分离的EV71在VP1区全长基因序列上无明显差别;但是随着时间推移,核苷酸变异的不断积累,出现新亚型的概率也在不断增加,因此有必要加强分离株的监测以掌握EV71的变异趋势.     

2010年致白城市手足口病流行的肠道病毒71型基因特征

目的分析2010年致白城市手足口病流行的肠道病毒71型基因特征。方法对2010年白城市6株EV71分离株VP1编码区基因全长逆转录-聚合酶链反应扩增后进行序列测定,使用Bioedit软件和MEGA 4.0软件进行同源性、基因亲缘关系分析。结果白城市2010年的6株EV71分离株均属于C4a基因亚型,6株白城分离株间的核苷酸同源性为98.6%～100.0%,并形成2个传播链,传播链1的白城4株病毒与吉林省2010年流行株代表株JL-HFM-10-5病毒亲缘关系最近,核苷酸同源性99.7%～100.0%,传播链2的2株白城流行株与2008年的阜阳流行株和2009年的2株吉林省流行株亲缘关系最近,核苷酸同源性99.3%～99.4%。结论 2010年白城市至少有两个EV71传播链致手足口病流行,传播链1的病毒在吉林省内和白城部分县区循环传播导致手足口病的流行;传播链2的病毒已经在国内传播较长时间,提示EV71病毒跨地域迅速而广泛传播特点。