2014-2016年天津市致病毒性脑炎人类肠道病毒基因特征分析
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摘要
目的了解天津市引起病毒性脑炎的人类肠道病毒(HEV)构成及其基因特征。方法收集2014-2016年天津市病毒性脑炎患者脑脊液和血清标本,通过细胞培养法分离病毒,采用RT-PCR分段扩增HEV VP1区全长序列并测序,进行同源性分析和基于VP1完整编码区的系统进化分析。结果从82份HEV阳性标本中分离到13株HEVB病毒,分离率15.85%,其中8株CV-B5型,2株E-6型,CV-B3型、E-18型、E-30型各1株。系统进化分析,天津市8株CV-B5与国内流行株C基因型亲缘关系较近,且分为两个不同的分支;HEV-B其它血清型均与国内流行株亲缘关系较近。结论天津市致病毒性脑炎HEV病原谱呈多样性特点,其中CV-B5为优势病原体,属于C基因型,且可能存在两条不同的传播链。
Objective To study the spectrum of human enteroviruses(HEV)causing viral encephalitis in Tianjin and their genetic characteristics. Methods Cerebrospinal fluid and serum samples were collected from patients with viral encephalitis in Tianjin from 2014-2016.Real-time RT-PCR was used to screen for HEV.Viruses were isolated via cell culture.The whole VP1 gene was then subjected to RT-PCR amplification,sequencing,homogeneity and phylogenetic tree analysis. Results Thirteen HEV-B strains were isolated from 82 samples at a rate of 15.85%.Eight of the isolates were CV-B5 strains,2 were E-6,1 was CV-B3,1 was E-18,and 1 was E-30.Phylogenetic tree analysis indicated that the genetic traits of the 8 CV-B5 strains isolated from Tianjin,which can be divided into two clusters,were close to those of genotype C strains in China.The two E-6 strains isolated from Tianjin were genotype D.The CV-B3 strains were genotype E,the E-18 strain was genotype C,and the E-30 strain was genotype II.All of the Tianjin HEV-B strains occupied the same clusters as the prevalent strains in China. Conclusion A vast spectrum of pathogens has caused viral encephalitis in Tianjin,and CV-B5 is the most prevalent of those pathogens.All of the CV-B5 strains from Tianjin were genotype C and may have two different chains of transmission.
Objective To study the spectrum of human enteroviruses(HEV)causing viral encephalitis in Tianjin and their genetic characteristics. Methods Cerebrospinal fluid and serum samples were collected from patients with viral encephalitis in Tianjin from 2014-2016.Real-time RT-PCR was used to screen for HEV.Viruses were isolated via cell culture.The whole VP1 gene was then subjected to RT-PCR amplification,sequencing,homogeneity and phylogenetic tree analysis. Results Thirteen HEV-B strains were isolated from 82 samples at a rate of 15.85%.Eight of the isolates were CV-B5 strains,2 were E-6,1 was CV-B3,1 was E-18,and 1 was E-30.Phylogenetic tree analysis indicated that the genetic traits of the 8 CV-B5 strains isolated from Tianjin,which can be divided into two clusters,were close to those of genotype C strains in China.The two E-6 strains isolated from Tianjin were genotype D.The CV-B3 strains were genotype E,the E-18 strain was genotype C,and the E-30 strain was genotype II.All of the Tianjin HEV-B strains occupied the same clusters as the prevalent strains in China. Conclusion A vast spectrum of pathogens has caused viral encephalitis in Tianjin,and CV-B5 is the most prevalent of those pathogens.All of the CV-B5 strains from Tianjin were genotype C and may have two different chains of transmission.
引文
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[14]李江,刘希佳,刘海舟,等.肠道病毒71型武汉分离株(EV71-WH029)全基因组cDNA克隆及序列分析[J].中国病原生物学杂志,2015,10(2):102-8.
[15]严菊英,卢亦愚,徐昌平,等.2002-2004年浙江省无菌性脑膜炎暴发疫情的病原学研究[J].病毒学报,2011,27(5):462-8.
[16]杨秀惠,严延生,翁育伟,等.福建省2011年病毒性脑炎暴发病原Echo30分子流行病学分析[J].中国人兽共患病学报,2012,28(3):265-9.
[2]Kennedy PGE,Quan PL,Lipkin WI.Viral encephalitis of unknown cause:current perspective and recent advances[J].Viruses,2017,9(6):pii:138.
[3]Chen X,Li J,Guo J,et al.An outbreak of echovirus 18encephalitis/meningitis in children in Hebei Province,China,2015[J].Emerg Microbes Infect,2017,6(6):e54.
[4]ZhuY,Zhou X,Liu J,et al.Molecular identification of human enteroviruses associated with aseptic meningitis in Yunnan Province,Southwest China[J].Springerplus,2016,5(1):1515.
[5]Yip CC,Lau SK,Woo PC,et al.Human enterovirus 71epidemics:what's next?[J].Emerg Health Threats J,2013(6):19780.
[6]徐国锋,王瑞,张清杰,等.河南省濮阳市7株肠道病毒71型全基因组序列分析[J].中国病原生物学杂志,2016,11(7):607-10,619.
[7]Oberste MS,Maher K,Klipatrick DR,et al.Molecular evolution of the human enteroviruses:correlation of serotype with VP1sequence and application to picornavirus classification[J].J Virol,1999,73(3):1941-8.
[8]Oberste MS,Maher K,Williams AJ,et al.Species-specific RTPCR amplification of human enteroviruses:a tool for rapid species identification of uncharacterized enteroviruses[J].J Gen Virol,2006,87(Pt 1):119-28.
[9]林小娟,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):143-7.
[10]韩文清,宫连凤,刘娟,等.烟台地区病毒性脑炎病毒分离与病原学分析[J].中国病原生物学杂志,2012,7(5):373-6.
[11]Chen P,Tao Z,Song Y,et al.A coxsackievirus B5-associated aseptic meningitis outbreak in Shandong Province,China in 2009[J].J Med Virol,2013,85(3):483-9.
[12]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):104-8.
[13]Tian H,Zhang Y,Sun Q,et al.Prevalence of multiple enteroviruses associated with hand,foot,and mouth disease in Shijiazhuang City,Hebei province,China:outbreaks of coxsackieviruses a10and b3[J].PLoS One,2014,9(1):e84233.
[14]李江,刘希佳,刘海舟,等.肠道病毒71型武汉分离株(EV71-WH029)全基因组cDNA克隆及序列分析[J].中国病原生物学杂志,2015,10(2):102-8.
[15]严菊英,卢亦愚,徐昌平,等.2002-2004年浙江省无菌性脑膜炎暴发疫情的病原学研究[J].病毒学报,2011,27(5):462-8.
[16]杨秀惠,严延生,翁育伟,等.福建省2011年病毒性脑炎暴发病原Echo30分子流行病学分析[J].中国人兽共患病学报,2012,28(3):265-9.