贵阳地区2013年手足口病非EV71、非CVA16型肠道病毒的检测分析
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摘要
目的了解贵阳地区引起手足口病(HFMD)的非肠道病毒71型(EV71)、非柯萨奇病毒A(CVA)16型肠道病毒的病原构成及优势型别,为贵阳地区HFMD防治工作提供依据。方法收集2013年4~6月检测为非EV71、非CVA16型肠道病毒的HFMD患者肛拭子标本共107例。提取病毒核酸,用巢式RT-PCR法扩增病毒VP4区序列,对PCR阳性扩增产物进行测序,通过与GenBank收录的序列BLAST比对确定病毒型别;并对优势型别进行序列和进化分析。结果 107例标本中共有100例标本巢式RT-PCR检测为阳性(93%),其中100例PCR产物测序成功,经BLAST比对后,100例标本病毒型别均得到确定:CVA6为46例,CVA10为30例,CVA5为10例,CVA2为6例,CVA3和CVA4各2例,CVB2和ECHO16各2例。CVA6在型别确定的非EV71、非CVA16型肠道病毒中占42.99%,为优势肠道病毒型别。结论贵阳地区引起HFMD的肠道病毒型别多样,CVA6为非EV71、非CVA16型肠道病毒中的优势型别。
Objective To explore the prevalence of non-EV71,non-CoxA16 enteroviruses strains of hand,foot and mouth disease(HFMD)in Guiyang,and analyze the genetic evolution of these non-EV71,non-CoxA16 enteroviruses strain,and to provide a basis for preventing and controlling(HFMD)in Guiyang.Methods A total of 107 HFMD specimens tested positive for enteroviruses and negative for EV71 and CoxA16 were collected from April to June of 2013.The virus RNA was extracted,and the VP4 sequence was amplified by nested RT-PCR.The PCR positive products were sequenced and the enteroviruses type was identified by sequence BLAST.The sequence and phylogenetic analysis of the predominant virus type was performed.Results 100 out of 107(93%)specimens were tested positive by nested RT-PCR,among which 100 PCR products were successfully sequenced.And the types of the 100 enterovirus strains were identifled by sequence BLAST:46CoxA6,30CoxA10,10CoxA5,6CoxA2,2CoxA3and2CoxA6,2CoxB2 and 2ECHO16.CoxA6 accounted for 42.99% in typed non-EV71,non-CoxA16 enterovimses,and was the predominant type.Conclusion Hand,foot and mouth disease in Guiyang was conceded with diverse enteroviruses,and CoxA6 was the predominant type among non-EV71,non-CoxA16 enteroviruses.
Objective To explore the prevalence of non-EV71,non-CoxA16 enteroviruses strains of hand,foot and mouth disease(HFMD)in Guiyang,and analyze the genetic evolution of these non-EV71,non-CoxA16 enteroviruses strain,and to provide a basis for preventing and controlling(HFMD)in Guiyang.Methods A total of 107 HFMD specimens tested positive for enteroviruses and negative for EV71 and CoxA16 were collected from April to June of 2013.The virus RNA was extracted,and the VP4 sequence was amplified by nested RT-PCR.The PCR positive products were sequenced and the enteroviruses type was identified by sequence BLAST.The sequence and phylogenetic analysis of the predominant virus type was performed.Results 100 out of 107(93%)specimens were tested positive by nested RT-PCR,among which 100 PCR products were successfully sequenced.And the types of the 100 enterovirus strains were identifled by sequence BLAST:46CoxA6,30CoxA10,10CoxA5,6CoxA2,2CoxA3and2CoxA6,2CoxB2 and 2ECHO16.CoxA6 accounted for 42.99% in typed non-EV71,non-CoxA16 enterovimses,and was the predominant type.Conclusion Hand,foot and mouth disease in Guiyang was conceded with diverse enteroviruses,and CoxA6 was the predominant type among non-EV71,non-CoxA16 enteroviruses.
引文
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[6]胡海赘,乔蓉,沈琦,等.2009~2011年上海市手足口病病原学检测及其临床特点分析[J].中华实验和临床病毒学杂志,2012,26(5):341-343.
[7]王彦霞,许汴利,陈豪敏,等.河南省2008~2010年手足口病流行趋势及病原学分析[J].中华预防医学杂志,2012,46(8):761-762.
[8]Blomqvist S,KlemoIa P,KaijaIairIen S,et al.C0-circulation of coxsackieviruses A6and A10in hand.foot and mouth disease out break in Finland[J].J Clin Virol,2010,48(1):49-54.
[9]Ang LW,Koh BK,Chart KP,et al.Epidemiology and Control of Hand,Foot and Mouth Disease in Singapore,2001-2007[J].Ann Acad Med Singapore,2009,38(2):106-112.
[10]Yamashita T,Ito M,Taniguchi A,et al.Prevalence of coxsackievirus A5,A6,and A10in patients with herpangina in Aichi Prefecture[J].Jpn J Infect Dis,2005,58(6):390-391.
[11]杨赫,陶泽新,王海岩,等.手足口病患儿柯萨奇病毒A10型山东地方株VPl区基因特征分析[J].中华传染病杂志,2010,28(7):385-389.
[2]Ishiko H,Shimada Y,Yonaha M,et al.Molecular diagnosis of human enteroviruses by phylogeny-based classification by use of the VP4sequence[J].J Infec Dis,2002,185(6):744-754.
[3]Kennett M,Birch C,Lewis F.Enterovirus type 71infection in Melbourne[J].Bull WHO,1974,51(6):609-615.
[4]杨兴林,熊金凤,李丽,等.贵阳地区2010年手足口病病原体检测分析[J].中国卫生检验杂志,2012,22(11):2734-2736.
[5]李静,金玉,檀晓娟,等.2010年南京市手足口病患儿的病原构成及肠道病毒71型分子流行病学特征分析[J].中华儿科杂志,2012,50(4):267-270.
[6]胡海赘,乔蓉,沈琦,等.2009~2011年上海市手足口病病原学检测及其临床特点分析[J].中华实验和临床病毒学杂志,2012,26(5):341-343.
[7]王彦霞,许汴利,陈豪敏,等.河南省2008~2010年手足口病流行趋势及病原学分析[J].中华预防医学杂志,2012,46(8):761-762.
[8]Blomqvist S,KlemoIa P,KaijaIairIen S,et al.C0-circulation of coxsackieviruses A6and A10in hand.foot and mouth disease out break in Finland[J].J Clin Virol,2010,48(1):49-54.
[9]Ang LW,Koh BK,Chart KP,et al.Epidemiology and Control of Hand,Foot and Mouth Disease in Singapore,2001-2007[J].Ann Acad Med Singapore,2009,38(2):106-112.
[10]Yamashita T,Ito M,Taniguchi A,et al.Prevalence of coxsackievirus A5,A6,and A10in patients with herpangina in Aichi Prefecture[J].Jpn J Infect Dis,2005,58(6):390-391.
[11]杨赫,陶泽新,王海岩,等.手足口病患儿柯萨奇病毒A10型山东地方株VPl区基因特征分析[J].中华传染病杂志,2010,28(7):385-389.