青岛市埃可病毒6型相关重症手足口病及疱疹性咽峡炎临床表现及病毒VP1区遗传进化分析
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摘要
目的了解青岛市埃可病毒6型(ECHO6)所致重症手足口病及疱疹性咽峡炎的临床特点及病毒基因特征。方法对2014年青岛市手足口病哨点1 727例手足口病及48例疱疹性咽峡炎患者的咽拭子标本进行肠道病毒检测,非EV-A71、非CV-A16咽拭子标本在RD及Hep-2细胞中进行毒株分离,半巢式反转录PCR扩增肠道病毒部分VP1序列,结合序列分析鉴定ECHO6毒株,对分离到的ECHO6毒株进行VP1编码区全长核苷酸序列测定,以Mega 7.0软件进行遗传进化及分子特征分析并分析相关病例的临床特征。结果共获得2株重症手足口病相关ECHO6毒株及1株疱疹性咽峡炎相关的ECHO6毒株VP1序列,2例手足口病重症病例均表现出高热、手、足、口腔部位皮疹和中枢神经系统受累症状。ECHO6青岛株VP1区核苷酸同源性为96.4%～97.6%,同属于D基因型D9亚型D9c分支,与2014年山东省无菌性脑膜炎病原中ECHO6毒株遗传关系密切。结论 ECHO6能引起中枢神经系统受累型重症手足口病,导致青岛市重症手足口病及疱疹性咽峡炎的ECHO6基因型属于D基因型D9亚型的D9c分支。
Objective To study the molecular characteristics of echovirus 6 and clinical manifestations of echovirus 6 associated severe hand,foot and mouth disease(HFMD) and herpangina in Qingdao city of China. Methods Throat swabs of 1 727 HFMD cases and 48 herpangina cases from HFMD sentinel at Qingdao city during 2014 were detected for enteroviruses(EVs).RD and hep-2 cells were used for virus proliferation; serotypes of CV-A16 were identified by partial VP1 gene sequencing with a semi-nested RT-PCR.The full-length of VP1 gene of 3 echovirus 6 strains(two isolates from severe HFMD and one from herpangina) were amplified and sequenced.Analysis was done using the MEGA7.0 software package.Clinical manifestation of echovirus 6 associated severe HFMD were also analyzed. Results Severe cases of ECHO6-related HFMD showed high fever, rashes on hands, feet, and oral sites, and symptoms of central nervous system involvement.The nucleotide homology of the VP1 region of all three virus strains was 96.4%-97.6%, which belong to the D9 c branch of the D genotype of ECHO6, and all showed close genetic relationship to ECHO6 strain isolated from aseptic meningitis in Shandong province from China at 2014. Conclusions ECHO6 is one of the pathogens of severe HFMD and herpangina in Qingdao city of China.D9c branch strains of the D9 subtype in D genotype are the major epidemic strains for severe HFMD and herpangina in recent years.
Objective To study the molecular characteristics of echovirus 6 and clinical manifestations of echovirus 6 associated severe hand,foot and mouth disease(HFMD) and herpangina in Qingdao city of China. Methods Throat swabs of 1 727 HFMD cases and 48 herpangina cases from HFMD sentinel at Qingdao city during 2014 were detected for enteroviruses(EVs).RD and hep-2 cells were used for virus proliferation; serotypes of CV-A16 were identified by partial VP1 gene sequencing with a semi-nested RT-PCR.The full-length of VP1 gene of 3 echovirus 6 strains(two isolates from severe HFMD and one from herpangina) were amplified and sequenced.Analysis was done using the MEGA7.0 software package.Clinical manifestation of echovirus 6 associated severe HFMD were also analyzed. Results Severe cases of ECHO6-related HFMD showed high fever, rashes on hands, feet, and oral sites, and symptoms of central nervous system involvement.The nucleotide homology of the VP1 region of all three virus strains was 96.4%-97.6%, which belong to the D9 c branch of the D genotype of ECHO6, and all showed close genetic relationship to ECHO6 strain isolated from aseptic meningitis in Shandong province from China at 2014. Conclusions ECHO6 is one of the pathogens of severe HFMD and herpangina in Qingdao city of China.D9c branch strains of the D9 subtype in D genotype are the major epidemic strains for severe HFMD and herpangina in recent years.
引文
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[16] Centers for Disease Control and Prevention.Nonpolio enterovirus and human parechovirus surveillance-United States,2006-2008[J].MMWR,2010,59(48):1577.
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[18] Cao CY,Chen QJ,He CR,et al.Analysis of genetic characteristics of ECHO6 virus isolated from an epidemic outbreak of encephalitis in Longyan,China[J].Bingdu Xuebao,2014,30(4):412-416.(in Chinese)曹春远,陈前进,何春荣,等.龙岩市一起暴发性脑炎疫情分离的埃可病毒 6 型基因特征分析[J].病毒学报,2014,30(4):412-416.
[19] Chen P,Lin X,Liu G,et al.Analysis of enterovirus types in patients with symptoms of aseptic meningitis in 2014 in Shandong,China[J].Virology,2018,516:196-201.
[20] Kim HJ,Kang B,Hwang S,et al.Epidemics of viral meningitis caused by echovirus 6 and 30 in Korea in 2008[J].Virol J,2012,9(1):38.
[21] Shi XY,Zhang YH,Su ZL,et al.Genetic characteristics of CV-A12 VP1 region and clinical manifestations of CV-A12-associated severe hand,foot and mouth disease in Qingdao[J].Zhonghua Weishengwuxue He Mianyixue Zazhi,2018,38(9):658-664.(in Chinese)史晓燕,张艳辉,苏志磊,等.青岛市柯萨奇病毒A12型VP1区基因特征及相关重症手足口病临床表现[J].中华微生物学和免疫学杂志,2018,38(9):658-664.
[22] Sun H,Huang X,Lin K,et al.Molecular evolution of two asymptomatic echovirus 6 strains that constitute a novel branch of recently epidemic echovirus 6 in China[J].Virol J,2017,14(1):140.
[2] Du XJ,Tao ZX,Liu GF,et al.Genetic characterization of echovirus 6 isolated from meningitis and encephalitis cases in Shandong Province,China[J].Bingdu Xuebao,2014,30(2):143-147.(in Chinese)杜小娟,陶泽新,刘桂芳,等.山东省脑膜炎和脑炎病例中埃可病毒6型的基因特征分析[J].病毒学报,2014,30(2):143-147.
[3] Li XL,Huang XY,Chen HM,et al.Genomic characteristics of echovirus 6 strains isolated from Henan province[J].Zhonghua Liuxingbingxue Zazhi,2012,33(8):876-877.(in Chinese)李幸乐,黄学勇,陈豪敏,等.河南省肠道病毒 Echo6 型脑炎病毒分离株全基因组序列分析[J].中华流行病学杂志,2012,33(8):876-877.
[4] Boell F,Meier-Ewert H.ECHO6-virus as the pathogen of hand,foot and mouth exanthema[J].Hautarzt ,1977,28(2):96-97.
[5] Nix WA,Oberste MS,Pallansch MA.Sensitive,seminested PCR amplification of VP1 sequences for direct identification of all enterovirus serotypes from original clinical specimens[J].J Clin Microbiol,2006,44(8):2698-2704.
[6] Mirand A,Archimbaud C,Henquell C,et al.Prospective identification of HEV-B enteroviruses during the 2005 outbreak[J].J Med Virol,2006,78(12):1624-1634.
[7] Pallansch MA,Oberste MS,Whitton JL.Enteroviruses:polioviruses,coxsackieviruses,echoviruses,and newer enteroviruses[M]//Knipe DM,Howley PM.Fields virology.Philadelphia:Lippincott Williams and Wilkins,2013:491-530.
[8] Lum LC,Chua KB,McMinn PC,et al.Echovirus 7 associated encephalomyelitis[J].J Clin Virol,2002,23(3):153-160.
[9] Danthanarayana N,Williams DT,Williams SH,et al.Acute meningoencephalitis associated with echovirus 9 infection in Sri Lanka,2009[J].J Med Virol,2015,87(12):2033-2039.
[10] Mladenova Z,Buttinelli G,Dikova A,et al.Aseptic meningitis outbreak caused by echovirus 30 in two regions in Bulgaria,May-August 2012[J].Epidemiol Infect,2014,142(10):2159-2165.
[11] Benschop KS,van der Avoort HG,Jusic E,et al.Polio and measles down the drain:environmental enterovirus surveillance in the Netherlands,2005 to 2015[J].Appl Environm Microbiol,2017,83(13):e00558-17.
[12] Wei XX,Zhang Y,Zhu SL,et al.Genetic characteristics of echovirus 6 isolated from the environmental sewage of Fengtai district in 2012,Beijing[J].Zhongguo Bingdubing Zazhi,2015,5(2):104-108.(in Chinese)尉秀霞,张勇,祝双利,等.北京市丰台区 2012 年污水中埃可病毒 6 型分离株 VP1 区的基因特征分析[J].中国病毒病杂志,2015,5(2):104-108.
[13] Jacques J,Moret H,Minette D,et al.Epidemiol ogical,molecular,and clinical features of enterovirus respiratory infections in French children between 1999 and 2005[J].J Clin Microbiol,2008,46(1):206-213.
[14] Suresh S,Forgie S,Robinson J.Non-polio enterovirus detection with acute flaccid paralysis:a systematic review[J].J Med Virol,2018,90(1):3-7.
[15] Messacar K,Abzug MJ,Dominguez SR.The changing epidemiology of acute flaccid paralysis warrants a paradigm shift in surveillance[J].J Med Virol,2018,90(1):1-2.
[16] Centers for Disease Control and Prevention.Nonpolio enterovirus and human parechovirus surveillance-United States,2006-2008[J].MMWR,2010,59(48):1577.
[17] Mao N,Zhao L,Zhu Z,et al.An aseptic meningitis outbreak caused by echovirus 6 in Anhui province,China[J].J Med Virol,2010,82(3):441-445.
[18] Cao CY,Chen QJ,He CR,et al.Analysis of genetic characteristics of ECHO6 virus isolated from an epidemic outbreak of encephalitis in Longyan,China[J].Bingdu Xuebao,2014,30(4):412-416.(in Chinese)曹春远,陈前进,何春荣,等.龙岩市一起暴发性脑炎疫情分离的埃可病毒 6 型基因特征分析[J].病毒学报,2014,30(4):412-416.
[19] Chen P,Lin X,Liu G,et al.Analysis of enterovirus types in patients with symptoms of aseptic meningitis in 2014 in Shandong,China[J].Virology,2018,516:196-201.
[20] Kim HJ,Kang B,Hwang S,et al.Epidemics of viral meningitis caused by echovirus 6 and 30 in Korea in 2008[J].Virol J,2012,9(1):38.
[21] Shi XY,Zhang YH,Su ZL,et al.Genetic characteristics of CV-A12 VP1 region and clinical manifestations of CV-A12-associated severe hand,foot and mouth disease in Qingdao[J].Zhonghua Weishengwuxue He Mianyixue Zazhi,2018,38(9):658-664.(in Chinese)史晓燕,张艳辉,苏志磊,等.青岛市柯萨奇病毒A12型VP1区基因特征及相关重症手足口病临床表现[J].中华微生物学和免疫学杂志,2018,38(9):658-664.
[22] Sun H,Huang X,Lin K,et al.Molecular evolution of two asymptomatic echovirus 6 strains that constitute a novel branch of recently epidemic echovirus 6 in China[J].Virol J,2017,14(1):140.