郭春颖,朱汝南,孙宇,赵林清,邓洁,王芳,宋秦伟,田润,钱渊

• 1:首都儿科研究所病毒研究室儿童病毒病病原学北京市重点实验室

摘要(Abstract)：

人偏肺病毒(human metapneumovirus,HMPV)是婴幼儿呼吸道感染的重要病原之一。本研究目的在于分析在HMPV监测中发现的少见型别A1基因亚型BJ-1610的全基因组序列,为HMPV的进一步研究提供数据支持。BJ-1610来源于一名患有肺炎的3月龄女婴的鼻咽洗液标本。对该毒株进行全基因组RNA分段扩增、测序、序列拼接并分析其基因组特征、变异程度及是否存在重组等现象。BJ-1610株基因组全长为13 406nt(KU821121)。BJ-1610与HMPV参考株进行全基因和各蛋白核苷酸和氨基酸序列比较,显示全基因组的核苷酸相似性范围为81.2%~98.4%,其中相似性最高的为澳大利亚株(KC562226),而SH和G基因则与其他澳大利亚株相似性更高。系统进化分析发现BJ-1610与HMPV A1基因亚型参考株位于同一分支,显示了与相似性分析相同的特征。对BJ-1610的膜表面蛋白F、SH和G蛋白序列进行分析结果显示,F蛋白最为保守,SH基因编码区的终止密码发生了突变,导致终止密码后移15位,长度由552bp变为567bp;G蛋白氨基酸序列突变较多,并导致N-糖基化位点的减少。HMPV作为儿童呼吸道疾病的重要病原之一,对其流行株基因组的分析将对其流行特征、遗传进化、致病机理、疫苗研制和抗病毒药物研发等方面的研究产生积极的推动作用。

关键词(KeyWords)： 儿童肺炎;人偏肺病毒;A1基因亚型;基因组;序列分析

Abstract:

Keywords:

基金项目(Foundation): 北京市卫生系统高层次卫生技术人才培养计划(项目号:2013-3-087);; 北京市科技计划(项目号:Z111107056811041),题目:北京儿童急性传染病监测体系的建立;; 国家自然科学基金(项目号:30872153),题目:新发现的病毒与儿科呼吸道感染的病原学研究

作者(Author): 郭春颖,朱汝南,孙宇,赵林清,邓洁,王芳,宋秦伟,田润,钱渊

DOI: 10.13242/j.cnki.bingduxuebao.003074

参考文献(References)：

• [1]van den Hoogen B G,de Jong J C,Groen J,Kuiken T,de Groot R,Fouchier R A,Osterhaus A D.A newly discovered human pneumovirus isolated from young children with respiratory tract disease[J].Nat Med,2001,7(6):719-724.
• [2]朱汝南,钱渊,邓洁,王芳,胡爱中,卢竞,曹力,袁艺,陈慧中.北京地区六岁以下儿童急性呼吸道偏肺病毒感染[J].中华儿科杂志,2003,41(6):441-444.
• [3]Boivin G,Abed Y,Pelletier G,Ruel L,Moisan D,CtéS,Peret T C,Erdman D D,Anderson L J.Virological features and clinical manifestations associated with human metapneumovirus:a new paramyxovirus responsible for acute respiratory-tract infections in all age groups[J].J Infect Dis,2002,186(9):1330-1334.
• [4]van den Hoogen B G,van Doornum G J,Fockens J C,Cornelissen J J,Beyer W E,de Groot R,Osterhaus A D,Fouchier R A.Prevalence and clinical symptoms of human metapneumovirus infection in hospitalized patients[J].J Infect Dis,2003,188(10):1571-1577.
• [5]van den Hoogen B G,Bestebroer T M,Osterhaus A D,Fouchier R A.Analysis of the genomic sequence of a human metapneumovirus[J].Virology,2002,295(1):119-132.
• [6]van den Hoogen B G,Herfst S,Sprong L,Cane P A,Forleo-Neto E,de Swart R L,Osterhaus A D,Fouchier R A.Antigenic and genetic variability of human metapneumoviruses[J].Emerg Infect Dis,2004,10(4):658-666.
• [7]Huck B,Scharf G,Neumann-Haefelin D,Puppe W,Weigl J,Falcone V.Novel human metapneumovirus sublineage[J].Emerg Infect Dis,2006,12(1):147-150.
• [8]Aberle J H,Aberle S W,Redlberger-Fritz M,Sandhofer M J,Popow-Kraupp T.Human metapneumovirus subgroup changes and seasonality during epidemics[J].Pediatr Infect Dis J,2010,29(11):1016-1018.
• [9]Zhu R N,Qian Y,Zhao L Q,Deng J,Sun Y,Wang F,Liao B,Li Y,Huang R Y.Characterization of human metapneumovirus from pediatric patients with acute respiratory infections in a 4-year period in Beijing,China[J].Chin Med J(Engl),2011,124(11):1623-1628.
• [10]Kong W,Wang Y,Zhu H,Lin X,Yu B,Hu Q,Yang X,Guo D,Peng J,Zhou D.Circulation of human metapneumovirus among children with influenza-like illness in Wuhan,China[J].Journal of Medical Virology,2016,88(5):774-781.
• [11]Zeng S,Xiao N,Zhong L,Yu T,Zhang B,Duan Z J.Clinical features of human metapneumovirus genotypes in children with acute lower respiratory tract infection in Changsha,China[J].Journal of Medical Virology,2015,87(11):1839-1845.
• [12]Ren J,Phan T,Bao X.Recent vaccine development for human metapneumovirus[J].J Gen Virol,2015,96(Pt7):1515-1520.
• [13]Zhu R N,Qian Y,Zhao L Q,Deng J,Sun Y,Wang F,Liao B,Li Y,Huang R Y.Characterization of human metapneumovirus from pediatric patients with acute respiratory infections in a 4-year period in Beijing,China[J].Chin Med J(Engl),2011,124(11):1623-1628.
• [14]Lo P A,Cammarota R,Apostoli P,Cella E,Fiorentini S,Babakir-Mina M,Ciotti M,Ciccozzi M.Genetic variability and circulation pattern of human metapneumovirus isolated in Italy over five epidemic seasons[J].New Microbiol,2011,34(4):337-344.
• [15]王芳,朱汝南,钱渊,邓洁,孙宇,赵林清,廖斌,黄蓉妍.人偏肺病毒基因型双重逆转录PCR检测方法的建立[J].中华检验医学杂志,2010,33(1):33-36.
• [16]Edwards K M,Zhu Y,Griffin M R,Weinberg G A,Hall C B,Szilagyi P G,Staat M A,Iwane M,Prill M M,Williams J V;New Vaccine Surveillance Network.Burden of human metapneumovirus infection in young children[J].N Engl J Med,2013,368(7):633-643.
• [17]朱汝南,钱渊,赵林清,邓洁,王芳,孙宇,廖斌.北京地区2004~2006年婴幼儿急性呼吸道感染中人偏肺病毒感染的研究[J].中华流行病学杂志,2007,28(7):679-682.
• [18]Kim J I,Park S,Lee I,Park K S,Kwak E J,Moon K M,Lee C K,Bae J Y,Park M S,Song K J.Genomewide analysis of human metapneumovirus evolution[J/OL].PLoS One,2016,11(4):e152962.
• [19]Piyaratna R,Tollefson S J,Williams J V.Genomic analysis of four human metapneumovirus prototypes[J].Virus Research,2011,160(1-2):200-205.
• [20]Cseke G,Maginnis M S,Cox R G,Tollefson S J,Podsiad A B,Wright D W,Dermody T S,Williams J V.Integrin alphavbeta1promotes infection by human metapneumovirus[J].Proc Natl Acad Sci USA,2009,106(5):1566-1571.
• [21]Cox R,Williams J.Breaking In:Human Metapneumovirus Fusion and Entry[J].Viruses,2013,5(1):192-210.
• [22]Bastien N,Normand S,Taylor T,Ward D,Peret T C,Boivin G,Anderson L J,Li Y.Sequence analysis of the N,P,M and F genes of Canadian human metapneumovirus strains[J].Virus Res,2003,93(1):51-62.
• [23]Wei Y,Zhang Y,Cai H,Mirza A M,Iorio R M,Peeples M E,Niewiesk S,Li J.Roles of the putative integrin-binding motif of the human metapneumovirus fusion(f)protein in cell-cell fusion,viral infectivity,and pathogenesis[J].J Virol,2014,88(8):4338-4352.
• [24]Bukreyev A,Whitehead S S,Murphy B R,Collins P L.Recombinant respiratory syncytial virus from which the entire SH gene has been deleted grows efficiently in cell culture and exhibits site-specific attenuation in the respiratory tract of the mouse[J].J Virol,1997,71(12):8973-8982.
• [25]Masante C,El Najjar F,Chang A,Jones A,Moncman C L,Dutch R E.The human metapneumovirus small hydrophobic protein has properties consistent with those of a viroporin and can Modulate viral fusogenic activity[J].Journal Virol,2014,88(11):6423-6433.
• [26]Thammawat S,Sadlon T A,Hallsworth P G,Gordon D L.Role of cellular glycosaminoglycans and charged regions of viral G protein in human metapneumovirus infection[J].Journal Virol,2008,82(23):11767-11774.
• [27]Skiadopoulos M H,Biacchesi S,Buchholz U J,Amaro-Carambot E,Surman S R,Collins P L,Murphy B R.Individual contributions of the human metapneumovirus F,G,and SH surface glycoproteins to the induction of neutralizing antibodies and protective immunity[J].Virology,2006,345(2):492-501.
• [28]Yang C,Wang C K,Tollefson S J,Lintao L D,Liem A,Chu M,Williams J V.Human metapneumovirus G protein is highly conserved within but not between genetic lineages[J].Archives of Virology,2013,158(6):1245-1252.