重庆地区儿童呼吸道合胞病毒和偏肺病毒分子流行病学研究
|
摘要
第一部分重庆地区急性呼吸道感染患儿呼吸道合胞病毒流行特征
目的分析重庆地区2008-2009年度急性呼吸道感染(ARTIs)住院患儿呼吸道合胞病毒的亚型流行情况,并了解优势流行株BA株的G蛋白基因特征。
方法采集2008年4月-2009年3月全年于重庆医科大学附属儿童医院因急性呼吸道感染住院的508例患儿鼻咽深部分泌物,用RT-PCR方法检测RSV并进行亚型鉴定,选取29例B亚型和10例A亚型RSV阳性标本,用RT-PCR的方法扩增全长G蛋白基因并测序。
结果在508例中,RSV阳性126例(24.8%),其中检测出A亚型43例(34.1%),B亚型80例(63.5%),A、B亚型混合感染3例(2.4%)。所测的10株A亚型的G基因与标准株A2的核苷酸同源性为91.4%～92%,均属GA2基因型;29株B亚型的G基因与标准株CH18537的核苷酸同源性为92.0%～93.0%,其中19株均为具有60个高度重复核苷酸插入的BA株。B亚型流行株与CH18537标准株相比,G基因有多种核苷酸变异如缺失、插入等,尤其在G蛋白近C端1/3处的高变区。
结论2008～2009年RSV仍是重庆地区儿童急性呼吸道感染的主要病原,与既往两年A亚型优势流行不同,2008-2009年度B亚型毒株流行占优;近年新发现的BA株可能已成为本地区优势流行株,BA株G基因变异是否导致G蛋白功能增强,进而促进其优势流行尚有待研究。
第二部分重庆地区急性呼吸道感染患儿人偏肺病毒流行特征
目的了解重庆地区2008-2009年度急性呼吸道感染(ARTIs)住院患儿人偏肺病毒的流行情况,并对hMPV的G基因序列进行比较分析。
方法收集2008年4月-2009年3月全年于重庆医科大学附属儿童医院因急性呼吸道感染住院的508例患儿鼻咽深部分泌物,用实时荧光定量PCR方法检测hMPV,再随机选取12例hMPV阳性标本,用RT-PCR的方法扩增全长G蛋白基因并测序,最后通过DNAStar软件对基因序列进行分析。
结果在508份标本中,hMPV检测阳性例数为60例,阳性率为11.8%。2008年4月至2009年3月间几乎每个月份(除2008年8月和9月)均有hMPV感染的阳性标本被检出,2008年4月至2008年6月为流行高峰,且2008年4月为最高峰,阳性率为31.9%。60例hMPV感染的临床标本中有9例标本存在与其他呼吸道病毒的协同感染。hMPV与同期的RSV的感染高峰没有重叠。hMPV感染患儿中5岁及以下患儿56例,占93.3%;其中2岁以下的患儿46例,占76.6% ;而5岁以上患儿仅有4例,占6.7%。hMPV感染的患儿与RSV感染后的临床症状类似,主要以肺炎和毛细支气管炎未主。12份hMPV阳性标本扩增的G基因进化树分析显示12株hMPV分为2种基因型和3种基因亚型,以CAN97-83为代表的基因型为主,其中所测的11株A亚型与CAN97-83的核苷酸同源性为82.6%～91.7%,氨基酸同源性69.5%～86.4%。另外一株B亚型与CAN98-75的核苷酸同源型为82.4%,氨基酸同源性为67.8%。
结论hMPV是重庆地区婴幼儿急性呼吸道感染(尤其是下呼吸道感染)的重要病毒病原之一,其感染对2岁以下婴幼儿威胁更大。重庆地区儿童感染的hMPV同时存在2种不同基因型,以CAN97-83为代表的A2基因型为主。
PARTⅠ:epidemiology features of respiratory syncytial virus in children with acute respiratory tract infections in Chongqing area
Objective To investigate epidemiological features of respiratory syncytial virus (RSV) A and B in hospitalized children with acute respiratory tract infections (ARTIs) and to analyze the genetic characteristics of G protein gene of RSV in Chongqing area, especially for BA stians.
Methods Nasopharyngeal secretions collected from 508 hospitalized children with ARTIs from April, 2008 to March,2009 were screened for RSV using RT-PCR. Full length G protein gene was amplified by RT-PCR from 10 RSV subtype A and 29 RSV subtype B strains.
Results Out of the total 508 specimens ,126(24.8%) were revealed positive for RSV. RSV subtype A and B viruses accounted for 34.1% and 63.5% of the total positive specimens, respectively. The remaining 2.4% of the specimens were positive for both subtype A and B. At the nucleotide level, identities between the 10 subtype A virus G genes and that of the prototype strain A2 were 91.4%～92%, indicating genotype GA2. Identities of the 29 subtype B virus G genes and that of the CH18537 strain were 92.0%～93.0%. Nineteen out of 29 RSV subtype B isolates contained highly repeated 60 nucleotides insertion in G protein gene, namely BA strain. As compared to the prototypes, the RSV G protein gene included nucleotide deletion, insertion, substitutions, especially in the carboxy-terminal third of the G gene.
Conclusion RSV has been the major cause of acute respiratory tract infections in children in Chongqing area. Subtype B strains, especially BA strains, were predominant during the study period. Whether the predominated circulation of BA strain is resulted from enhanced attachment function of G protein remains unknown.
PARTⅡ:epidemiology features of human metapneumovirus in children with acute respiratory tract infections in Chongqing area
Objective To investigate epidemiological features of human metapneumovirus (hMPV) in hospitalized children with acute respiratory tract infections (ARTIs) and to analyze the genetic characteristics of G protein gene of hMPV in Chongqing area.
Methods Nasopharyngeal secretions collected from 508 hospitalized children with ARTIs from April, 2008 to March,2009 were screened for hMPV using real-time reverse transcription-PCR. Full length G protein gene was amplified by RT-PCR from 12 strains hMPV.
Results During the study period,out of the total 508 clinical specimens,60(11.8%) were revealed positive for hMPV.hMPV was detected almost in every month from April 2008 to March 2009 (except August and september,2008). A peak was noticed in April 2008 to July 2008, especially April 2008. Among 60 positive samples, 9 were also positive for other respiratory viruses,suggesting that these children were co-infected with hMPV and other respiratory viruses. The peak of hMPV prevalence did not overlap with that of RSV during this period. Fifty-six out of sixty positive (93.3%,56/60) specimens on hMPV were from children under 5 year olds, forty-six (76.6%,46/60) were younger than 2 years of age, and four (6.7%,4/60) were over 5 years of age. The clinical manifestations of hMPV and RSV infections had a similar symptoms. Phylogenetic tree analysis of nucleotide sequences revealed two different genotype and three subtypes circulating during the period. At the nucleotide level, identities between the 11 subtype A virus G gene and that of the prototype strain CAN97-83 were 82.6%～91.7%, and the amino acid identities were 69.5%～86.4%. The nucleotide identities between the one subtype B virus G gene and that of the CAN98-75 was 82.4% and the amino acid identities was 67.8%.
Conclusion our findings suggested that hMPV had been one of the important agents causing acute respiratory infections in Chongqing area. Infants and young children under two years of age seemed to be more susceptible to hMPV. There were two different genotypes of hMPV in Chongqing area, and CAN97-83 was the dominant type in Chongqing area.
目的分析重庆地区2008-2009年度急性呼吸道感染(ARTIs)住院患儿呼吸道合胞病毒的亚型流行情况,并了解优势流行株BA株的G蛋白基因特征。
方法采集2008年4月-2009年3月全年于重庆医科大学附属儿童医院因急性呼吸道感染住院的508例患儿鼻咽深部分泌物,用RT-PCR方法检测RSV并进行亚型鉴定,选取29例B亚型和10例A亚型RSV阳性标本,用RT-PCR的方法扩增全长G蛋白基因并测序。
结果在508例中,RSV阳性126例(24.8%),其中检测出A亚型43例(34.1%),B亚型80例(63.5%),A、B亚型混合感染3例(2.4%)。所测的10株A亚型的G基因与标准株A2的核苷酸同源性为91.4%～92%,均属GA2基因型;29株B亚型的G基因与标准株CH18537的核苷酸同源性为92.0%～93.0%,其中19株均为具有60个高度重复核苷酸插入的BA株。B亚型流行株与CH18537标准株相比,G基因有多种核苷酸变异如缺失、插入等,尤其在G蛋白近C端1/3处的高变区。
结论2008～2009年RSV仍是重庆地区儿童急性呼吸道感染的主要病原,与既往两年A亚型优势流行不同,2008-2009年度B亚型毒株流行占优;近年新发现的BA株可能已成为本地区优势流行株,BA株G基因变异是否导致G蛋白功能增强,进而促进其优势流行尚有待研究。
第二部分重庆地区急性呼吸道感染患儿人偏肺病毒流行特征
目的了解重庆地区2008-2009年度急性呼吸道感染(ARTIs)住院患儿人偏肺病毒的流行情况,并对hMPV的G基因序列进行比较分析。
方法收集2008年4月-2009年3月全年于重庆医科大学附属儿童医院因急性呼吸道感染住院的508例患儿鼻咽深部分泌物,用实时荧光定量PCR方法检测hMPV,再随机选取12例hMPV阳性标本,用RT-PCR的方法扩增全长G蛋白基因并测序,最后通过DNAStar软件对基因序列进行分析。
结果在508份标本中,hMPV检测阳性例数为60例,阳性率为11.8%。2008年4月至2009年3月间几乎每个月份(除2008年8月和9月)均有hMPV感染的阳性标本被检出,2008年4月至2008年6月为流行高峰,且2008年4月为最高峰,阳性率为31.9%。60例hMPV感染的临床标本中有9例标本存在与其他呼吸道病毒的协同感染。hMPV与同期的RSV的感染高峰没有重叠。hMPV感染患儿中5岁及以下患儿56例,占93.3%;其中2岁以下的患儿46例,占76.6% ;而5岁以上患儿仅有4例,占6.7%。hMPV感染的患儿与RSV感染后的临床症状类似,主要以肺炎和毛细支气管炎未主。12份hMPV阳性标本扩增的G基因进化树分析显示12株hMPV分为2种基因型和3种基因亚型,以CAN97-83为代表的基因型为主,其中所测的11株A亚型与CAN97-83的核苷酸同源性为82.6%～91.7%,氨基酸同源性69.5%～86.4%。另外一株B亚型与CAN98-75的核苷酸同源型为82.4%,氨基酸同源性为67.8%。
结论hMPV是重庆地区婴幼儿急性呼吸道感染(尤其是下呼吸道感染)的重要病毒病原之一,其感染对2岁以下婴幼儿威胁更大。重庆地区儿童感染的hMPV同时存在2种不同基因型,以CAN97-83为代表的A2基因型为主。
PARTⅠ:epidemiology features of respiratory syncytial virus in children with acute respiratory tract infections in Chongqing area
Objective To investigate epidemiological features of respiratory syncytial virus (RSV) A and B in hospitalized children with acute respiratory tract infections (ARTIs) and to analyze the genetic characteristics of G protein gene of RSV in Chongqing area, especially for BA stians.
Methods Nasopharyngeal secretions collected from 508 hospitalized children with ARTIs from April, 2008 to March,2009 were screened for RSV using RT-PCR. Full length G protein gene was amplified by RT-PCR from 10 RSV subtype A and 29 RSV subtype B strains.
Results Out of the total 508 specimens ,126(24.8%) were revealed positive for RSV. RSV subtype A and B viruses accounted for 34.1% and 63.5% of the total positive specimens, respectively. The remaining 2.4% of the specimens were positive for both subtype A and B. At the nucleotide level, identities between the 10 subtype A virus G genes and that of the prototype strain A2 were 91.4%～92%, indicating genotype GA2. Identities of the 29 subtype B virus G genes and that of the CH18537 strain were 92.0%～93.0%. Nineteen out of 29 RSV subtype B isolates contained highly repeated 60 nucleotides insertion in G protein gene, namely BA strain. As compared to the prototypes, the RSV G protein gene included nucleotide deletion, insertion, substitutions, especially in the carboxy-terminal third of the G gene.
Conclusion RSV has been the major cause of acute respiratory tract infections in children in Chongqing area. Subtype B strains, especially BA strains, were predominant during the study period. Whether the predominated circulation of BA strain is resulted from enhanced attachment function of G protein remains unknown.
PARTⅡ:epidemiology features of human metapneumovirus in children with acute respiratory tract infections in Chongqing area
Objective To investigate epidemiological features of human metapneumovirus (hMPV) in hospitalized children with acute respiratory tract infections (ARTIs) and to analyze the genetic characteristics of G protein gene of hMPV in Chongqing area.
Methods Nasopharyngeal secretions collected from 508 hospitalized children with ARTIs from April, 2008 to March,2009 were screened for hMPV using real-time reverse transcription-PCR. Full length G protein gene was amplified by RT-PCR from 12 strains hMPV.
Results During the study period,out of the total 508 clinical specimens,60(11.8%) were revealed positive for hMPV.hMPV was detected almost in every month from April 2008 to March 2009 (except August and september,2008). A peak was noticed in April 2008 to July 2008, especially April 2008. Among 60 positive samples, 9 were also positive for other respiratory viruses,suggesting that these children were co-infected with hMPV and other respiratory viruses. The peak of hMPV prevalence did not overlap with that of RSV during this period. Fifty-six out of sixty positive (93.3%,56/60) specimens on hMPV were from children under 5 year olds, forty-six (76.6%,46/60) were younger than 2 years of age, and four (6.7%,4/60) were over 5 years of age. The clinical manifestations of hMPV and RSV infections had a similar symptoms. Phylogenetic tree analysis of nucleotide sequences revealed two different genotype and three subtypes circulating during the period. At the nucleotide level, identities between the 11 subtype A virus G gene and that of the prototype strain CAN97-83 were 82.6%～91.7%, and the amino acid identities were 69.5%～86.4%. The nucleotide identities between the one subtype B virus G gene and that of the CAN98-75 was 82.4% and the amino acid identities was 67.8%.
Conclusion our findings suggested that hMPV had been one of the important agents causing acute respiratory infections in Chongqing area. Infants and young children under two years of age seemed to be more susceptible to hMPV. There were two different genotypes of hMPV in Chongqing area, and CAN97-83 was the dominant type in Chongqing area.
引文
[1] Kap lan NM, DoveW, Abd-Eldayem SA, et al. Molecular epidemiology and disease severity of respiratory syncytial virus in relation to other potentialpathogens in children hosp italized with acute respiratory infection in Jordan[J]. J Med Virol, 2008, 80 (1) : 168-174.
[2] Yong S, O’Keefe PTO, Amott J, et al. Lungfunction airway responsiveness and respiratory symptoms before and after bronchilitis[J]. Arch Dis Child. 1995,72;16.
[3] Hacking D,Hull J. Respiratory syncytial virus-viral biology and the host response [J].J Infect, 2002 Jul,45(1);18-24.
[4] Johnson PR, Spriggs MK, Olmsted RA, et al. The G glycoprotein of human respiratory syncytial virus of subgroup A and B extensive sequence divergence between antigenically related proteins[J].Proc Natl Acad Sci USA, 1987, 84: 5625- 5629.
[5] Hall CB, Waish EE, Schnabel KC,et al. Occurrence of groups A and B of respiratory syncytialvirus over 15 years: associated epidemiologic and clinical characteristics in hospitalized and ambulatory children[J]. J Inf Disea, 1990,162: 1283-1290.
[6] Chan PKS, Sung RYT, Fung KSC, et al. Ep idemiology of respiratory syncytial virus infection among pediatric patients in Hong Kong: seasonality and disease impact [J]. J Ep idemiol Infect,1999, 123: 2572262.
[7]朱汝南,耿学辉,王之栋,等.应用逆转录-聚合酶链反应鉴别呼吸道合胞病毒亚型[J].中华儿科杂志,1998,36:538-540.
[8] Rajala MS, Sullender WM,Prasad AK, et al. Genetic variability among Group A and B respiratory syncytial virus isolates from a large referral hospital in New Delhi[J], India.J Clin Microbial, 2003, 41:2311-2316.
[9] Trento A, Viegas M, Galiano, et al. Natural history of human respiratory syncytial virus inferred from phylogenetic analysis of the attachment (G) glycoprotein with a 60-nucleotide duplication[J]. J Viro,2006, 80. 975-984.
[10] Venkatesh MP, Weisman LE.Prevent and treatment of respiratory syncytial virus infection infants:an update[J]. Expert Rev Vaccines, 2006, 5:261-268.
[11] Sonoda S, Getoh Y, Bann F, et al. Acute lower respiratory infections inhospitalized children over a 6 year period in tokyo[J]. Pediatr Int.1999, 41:519- 524.
[12]张志勇,陈昕,赵晓东,等.急性呼吸道感染儿童两亚型呼吸道合胞病毒检测分析[J].中国实用儿科杂志,2008,23:905-908.
[13]邓洁,钱渊,朱汝南,等.2000年冬-2006年春北京地区急性呼吸道感染患儿中呼吸道合胞病毒的监测[J].中华儿科杂志,2006,44:924-927.
[14] Peret TC,Hall CB,Schnabel KC,et al. Cirulation patterns of genetically distinct group A and B strains of human respiratory syncytial virus in a community[J]. J Gen virol,1998,79:2221-2229.
[15] Trento A, Galiano M, Videla C, et a1. Major changes in the G protein of human respiratory syncytial virus isolates introduced by a duplication of 60 nucleotides. [J] J Gen Virol,2003,84:3ll5-3120.
[16] Silva LHA, Spilki FR, Riccetto AGL, et al. Genetic Variability in the G Protein gene of human respiratory syncytial Virus isolated from the campinas metropolitan region, Brazil[J]. J Med Viro,2008, 80:1653-1660.
[17] Zlateva KT, lemey P, Mose E, et al. Genetic variability and molecular evolution of the human respiratory syncytial virus subgroup B attachment G protein[J]. J Viro, 2005,79:9157-9167.
[18]邓洁,朱汝南,钱渊,等.北京地区2000-2004年分离的呼吸道合胞病毒B亚型株G蛋白基因的序列分析[J].中华微生物学和免疫学杂志,2006,26: 1-5.
[19] Ball LA. Replication strategies of RNA viruses. Fields Virology. Philadelphia :Lippincott Willams&Wikins.pp105-108.
[20]耿学辉,王之梁,钱渊,等.呼吸道合胞病毒G蛋白的基因分析[J].病毒学报[J],1999,39:39-41.
[1] van den Hoogen BG,de Jong JC,Groen J,et al.A newly discovered human pneumovirusisolated from young children with respiratory tract disease[J].Nat Med,2001 ,7(6):719-724.
[2] Chan PKS, Tam JS, Lam C-W, et al. Human metapneumovirus detection in patients with severe acute respiratory syndrome [J]. Emerg Infect Dis, 2003;9(9):1058-1063.
[3] Frank Esper, MD, Derek Boucher, et al. Human Metapneumovirus Infection in the United States: Clinical Manifestations Associated With a Newly Emerging Respiratory Infection in Children [J]. Pediatrics, 2003, 6(111) : 1407-1410.
[4] Nathalie Bastien, Diane Ward, Paul Van Caeseele, et al. Human Metapneumovirus Infection in the Canadian Population [J]. J. Clin. Microbiol., 2003; 10(41):4642- 4646.
[5] van den Hoogen, B. G., T. M. Bestebroer, et al. Analysis of the genomic sequence of a human metapneumovirus[J]. Virology, 2002; 295:119-132.
[6] Boivin G, Mackay I, Sloots TP, et al. Global genetic diversity of humanmetapneumovirus fusion gene [J]. Emerg Infect Dis, 2004; 10:1154–1157.
[7] Biacchesi S, Skiadopoulos MH, Boivin G, et al. Genetic diversity between human metapneumovirus subgroups[J]. Virology,2003; 315:1–9.
[8] Gregory CG, Ana WC, Sharon F, et al. Multi-year study of human metapneumovirus infection at a large US midwestern medical referral center [J]. J Clin Virol. 2006; 37(4):269-276.
[9] Murray CJL, Lopez AD, Mathers CD, et al. The global burden of disease 2000 project : aim, methods and data sources. Global programme on evidence for health policy.Geneva: World Health org, 2001,136.
[10] Ordas J, Boga JA, Alvarez-Arguelles M, et a1. Role of metapneumovlrus in viral respiratory infections in young children [J]. J Clin Microbiol 2006, 44(8): 2739-2742
[11] Mackay IM, Bialasiewicz S, Waliuzzaman Z, et a1. Use of the P gene to genotype human metapneumovirus identifies 4 viral subtypes [J] .J Infect Dis, 2004, 190 (11):1913-1918.
[12] Scheltinga SA, Templeton KE, Beemma MF, et a1. Diagnosis of human metapneumovirus and rhinovirus in patients with respiratory tract infections by an internally controlled multiplex real-time RNA PCR [J]. J Clin Virol, 2005, 33(4): 306-311.
[13] Sumino KC,Asapov E,Pierce RA , et a1.Detection of severe human metapneumovirus infection by real-time polymerase chain reaction and histopatho-logical assessment [J].J Infect Dis, 2005, 192(6):1052—1060.
[14]朱汝南,钱渊,赵林清,等.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.
[15]陈慧中,钱渊,王天有,等.偏肺病毒毛细支气管炎的临床研究[J].中华儿科杂志, 2004, 42(5):383-386.
[16] Peiris JS, TangWH, Chart KH, et al. Children with respiratory disease associated with metapneumovirus in HongKong[J].Emerg Infect Dis, 2003, 9(6): 628-633.
[17] van den Hoogen BG, Osterhaus DM, Fouchier RA. Clinical impact and diagnosis of human metapneumovirus infection[J].Pediatr Infect Dis J, 2004, 23:25-32
[18] van den Hoogen BG, van Doomum GJ, Fockem JC, et al. Prevalence and clinical symptoms of human metapneumovirus infecion in hospitalized patients[J]. J Infect Dis, 2003, 188:1571-1577.
[19] Esper F, Martinello RA, Boucher D, et al. A 1-year experience with human metapneumovirus in children aged<5 years[J]. J Infect Dis, 2004, 189: 1388-1396.
[20] Bastien N, Ward D, Van Caeseele PK, et al. Human metapneumovirus infection in the canadian population [J].J Clin Microbiol, 2003, 41(10): 4642-4646.
[21] Peiris JS, TangWH, Chart KH, et al. Children with respiratory disease associated with metapneumovirus in HongKong [J].Emerg Infect Dis, 2003, 9(6):628-633.
[22] Gerna G, Campanini G, Rovida F, et al. Changing circulation rate of human metapneumovirus stains and types among hospitalized pediatric patients during three consecutive winter-spring seasons[J]. Arch Virol, 2005, 150: 2365-2375.
[23] Greensill J,McNamara PS, Dove W, et al. Human metapneumovirus in sever respiratory syncytial virus bronchiolitis[J]. Emerg infect Dis 2003,9:372-375.
[24] kuypers J, Wright N, Corey L, et al. Detection and quantification of human metapneumovirus in pediatric specimens by real-time RT-PCR [J]. J Clin Virol 2005,33:299-305.
[25] Semple MG, Cowell A, Dove W, et al. Dual infection of infants by human metapneumovirus and human respiratory syncytial virus is strongly associated with severe bronchiolitis[J].J Infect Dis ,2005, 191:382-386.
[26] Nissen MD, Siebert DJ, Mackay IM, et al. Evidence of human metapneumovirus in Australian children[J]. Med J Aust 2002,176,:188.
[27] Boivin G, Abed Y, Pelletier G, et al.Virological features and clinicalmanifestations associated with human metapneumovirus:a new paramyxovirus responsible for acute respiratory-tract infections in all age groups[J].J.Infect.Dis. 2002, 186:1130-1334.
[1] van den Hoogen BG, deJong JC, Groen J, et al. A newly discovered human pneumovirus isolated from young children with respiratory tract disease [J]. Nat Med, 2001; 7: 719–724.
[2] Chan PKS, Tam JS, Lam C-W, et al. Human metapneumovirus detection in patients with severe acute respiratory syndrome [J]. Emerg Infect Dis, 2003;9(9):1058-1063.
[3] Frank Esper, MD, Derek Boucher, et al. Human Metapneumovirus Infection in the United States: Clinical Manifestations Associated With a Newly Emerging Respiratory Infection in Children [J]. Pediatrics,2003;6(111) : 1407-1410.
[4] Nathalie Bastien, Diane Ward, Paul Van Caeseele, et al. Human Metapneumovirus Infection in the Canadian Population [J]. J. Clin. Microbiol., 2003; 10(41):4642-4646.
[5] Biacchesi S, Skiadopoulos MH, Boivin G, et a1. Genetic diversity between human pneumovirus subground[J]. Virology. 2003, 315:1-9.
[6] van den Hoogen, B. G., T. M. Bestebroer, et al. Analysis of the genomic sequence of a human metapneumovirus[J]. Virology, 2002; 295:119-132.
[7] Boivin G, Abed Y, Pelletier G, et al.Virological features and clinical manifestations associated with human metapneumovirus:a new paramyxovirus responsible for acute respiratory-tract infections in all age group[J]s.J.Infect.DIS .2002, 186:1130-1334.
[8] Pelletier G, Dery P, Abed Y, Boivin G.Respiratory tract reinfections by the new human metapneumovirus in an immunocompromised child. Emerg . Infect . Dis . 2002, 8:976-978.
[9] Williams J, Harris P, Tollefson S, et al. Human metapneumovirus and lower respiratory tract disease in otherwise healthy infants and children, N. Engl . J . Med. 2004,350:443-450.
[10] van den Hoogen BG, Osterhaus DM, Fouchier RA. Clinical impact and diagnosis of human metapneumovirus infection[J].Pediatr Infect Dis J, 2004, 23: 25-32.
[11] van den Hoogen BG, van Doomum GJ, Fockem JC, et al. Prevalence and clinical symptoms of human metapneumovirus infecion in hospitalized patients[J]. J Infect Dis, 2003, 188:1571-1577.
[12] Esper F, Martinello RA, Boucher D, et al. A 1-year experience with human metapneumovirus in children aged<5 years[J]. J Infect Dis, 2004, 189: 1388-1396.
[13] Bastien N, Ward D, Van Caeseele PK, et al. Human metapneumovirus infection in the canadian population [J]. J Clin Microbiol, 2003, 41(10): 4642-4646.
[14] Peiris JS, TangWH, Chart KH, et al. Children with respiratory disease associated with metapneumovirus in HongKong[J].Emerg Infect Dis, 2003, 9(6):628-633.
[15]朱汝南,钱渊,邓洁,等.北京地区六岁以下儿童急性呼吸道偏肺病毒感染.中华儿科杂志[J],2003,41:441-444.
[16] Dollner H, Risnes K, Radtke A, et al. Outbreak of human metapneumovirus infection in Norwegian children[J]. Pediatr, J Infect Dis, 2004, 23:436-439.
[17] Mackay IM,Bialasiewicz S,Waliuzzaman Z, et a1.Use of the P gene to genotype human metapneumovirus identifies 4 viral subtypes [J] .J Infect Dis, 2004, 190 (11):1913-1918.
[18] Scheltinga SA, Templeton KE, Beemma MF, et a1.Diagnosis of human metapneumovirus and rhinovirus in patients with respiratory tract infections by an internally controlled multiplex real-time RNA PCR [J]. J Clin Virol, 2005, 33(4): 306-311.
[19] Sumino KC,Asapov E,Pierce RA , et a1.Detection of severe human metapneumovirus infection by real-time polymerase chain reaction and histopatho-logical assessment [J].J Infect Dis, 2005, 192(6):1052—1060.
[20] Louis JK,Hacker JK,Gonzales R,et a1.Characterization of viral agents causing acute respiratory infection in a San Francisco University Medical Center Clinic during the influenza season[J].Clin Infect Dis,2005,41(6):822-828.
[21] Noyola DE,Alpuche-Solis AG,Herrera-Diaz A,et a1.Human metapneum- ovirus infections in Mexico: epidemiological and clinical characteristics [J]. J Med Microbiol, 2005,54:969-974.
[22]陈慧中,钱渊,王天有,等.偏肺病毒毛细支气管炎的临床研究[J].中华儿科杂志,2004,42(5):383—386.
[23] Esper F, Boucher D, Weibel C, et a1. Human metapneumovirus infection in the United States:clinical manifestation associated with a newly emerging respiratory infection in children[J]. Pediatrics, 2003, 111:1407-1410.
[24] Rawlinson W,Waliuzzaman Z,Carter I ,et al.Asthma exacerbations in children associated with rhinovirus but no human metapneumovirus infection[J] J.Infect. Dis.2003,187:1314-1318.
[25] Williams J, Harris P, Tollefson S, et al. Human metapneumovirus and lower respiratory tract disease in otherwise healthy infants and children, N. Engl. J Med.2004,350:443-450.
[26] Carla P, van den Hoogen B, Chakraharti S, et al. Human metapneumovirus in a haematopoietic stem cell transplant recipient with fatal lower respiratory tractdiease[J]. Bone Marrow Transplant. 2003,31:309-310.
[27] Jartti T, Van den Hoogen B, Carofalo RP,et a1. Metapneumovirus and acute wheezing in children[J]. Lancet, 2002, 360(9343):1393-1394.
[28] Greensill J, McNamara P, Dove W , et al. Human metapneumovirus in severe respiratory syncytial virus bronchiolitis[J]. Emerg. Infect.Dis.2003,9 :372—375.
[29] Bosis S, Esposito S, Niesters H, et al. Impact of human metapneumovirus in childhood:Comparison with respiratory syncytial virus and influenza viruses[J]. J.Med virol .2005,75:101-104.
[30] Martinez I, Melero J. Binding of human respiratory syncytial virus to cells: implication of sulfated cell surface proteoglycans[J].J. Gen. Viro1.2000. 81: 2715-2722.
[31] Tripp R, Jones L, Haynes L, et al. CX3C chemokine mimicry by respiratory syncytial virus G glycoprotein. Nat. Immunol . 2001, 2:732-738.
[32] Peret T, Abed Y, Erdman D, et al. Sequence polymorphism of the predicted human metapneumovirus G glycoprotein [J].J. Gan. Viro1. 2004. 85:679-686.
[33] Alvarez R, Harrod K, Shieh W, et al. Human metapneumovirus persists in BALB/c mice despite the presence of neutralizing antibodies[J]. J.Virol. 2004. 78:14003-14011.
[2] Yong S, O’Keefe PTO, Amott J, et al. Lungfunction airway responsiveness and respiratory symptoms before and after bronchilitis[J]. Arch Dis Child. 1995,72;16.
[3] Hacking D,Hull J. Respiratory syncytial virus-viral biology and the host response [J].J Infect, 2002 Jul,45(1);18-24.
[4] Johnson PR, Spriggs MK, Olmsted RA, et al. The G glycoprotein of human respiratory syncytial virus of subgroup A and B extensive sequence divergence between antigenically related proteins[J].Proc Natl Acad Sci USA, 1987, 84: 5625- 5629.
[5] Hall CB, Waish EE, Schnabel KC,et al. Occurrence of groups A and B of respiratory syncytialvirus over 15 years: associated epidemiologic and clinical characteristics in hospitalized and ambulatory children[J]. J Inf Disea, 1990,162: 1283-1290.
[6] Chan PKS, Sung RYT, Fung KSC, et al. Ep idemiology of respiratory syncytial virus infection among pediatric patients in Hong Kong: seasonality and disease impact [J]. J Ep idemiol Infect,1999, 123: 2572262.
[7]朱汝南,耿学辉,王之栋,等.应用逆转录-聚合酶链反应鉴别呼吸道合胞病毒亚型[J].中华儿科杂志,1998,36:538-540.
[8] Rajala MS, Sullender WM,Prasad AK, et al. Genetic variability among Group A and B respiratory syncytial virus isolates from a large referral hospital in New Delhi[J], India.J Clin Microbial, 2003, 41:2311-2316.
[9] Trento A, Viegas M, Galiano, et al. Natural history of human respiratory syncytial virus inferred from phylogenetic analysis of the attachment (G) glycoprotein with a 60-nucleotide duplication[J]. J Viro,2006, 80. 975-984.
[10] Venkatesh MP, Weisman LE.Prevent and treatment of respiratory syncytial virus infection infants:an update[J]. Expert Rev Vaccines, 2006, 5:261-268.
[11] Sonoda S, Getoh Y, Bann F, et al. Acute lower respiratory infections inhospitalized children over a 6 year period in tokyo[J]. Pediatr Int.1999, 41:519- 524.
[12]张志勇,陈昕,赵晓东,等.急性呼吸道感染儿童两亚型呼吸道合胞病毒检测分析[J].中国实用儿科杂志,2008,23:905-908.
[13]邓洁,钱渊,朱汝南,等.2000年冬-2006年春北京地区急性呼吸道感染患儿中呼吸道合胞病毒的监测[J].中华儿科杂志,2006,44:924-927.
[14] Peret TC,Hall CB,Schnabel KC,et al. Cirulation patterns of genetically distinct group A and B strains of human respiratory syncytial virus in a community[J]. J Gen virol,1998,79:2221-2229.
[15] Trento A, Galiano M, Videla C, et a1. Major changes in the G protein of human respiratory syncytial virus isolates introduced by a duplication of 60 nucleotides. [J] J Gen Virol,2003,84:3ll5-3120.
[16] Silva LHA, Spilki FR, Riccetto AGL, et al. Genetic Variability in the G Protein gene of human respiratory syncytial Virus isolated from the campinas metropolitan region, Brazil[J]. J Med Viro,2008, 80:1653-1660.
[17] Zlateva KT, lemey P, Mose E, et al. Genetic variability and molecular evolution of the human respiratory syncytial virus subgroup B attachment G protein[J]. J Viro, 2005,79:9157-9167.
[18]邓洁,朱汝南,钱渊,等.北京地区2000-2004年分离的呼吸道合胞病毒B亚型株G蛋白基因的序列分析[J].中华微生物学和免疫学杂志,2006,26: 1-5.
[19] Ball LA. Replication strategies of RNA viruses. Fields Virology. Philadelphia :Lippincott Willams&Wikins.pp105-108.
[20]耿学辉,王之梁,钱渊,等.呼吸道合胞病毒G蛋白的基因分析[J].病毒学报[J],1999,39:39-41.
[1] van den Hoogen BG,de Jong JC,Groen J,et al.A newly discovered human pneumovirusisolated from young children with respiratory tract disease[J].Nat Med,2001 ,7(6):719-724.
[2] Chan PKS, Tam JS, Lam C-W, et al. Human metapneumovirus detection in patients with severe acute respiratory syndrome [J]. Emerg Infect Dis, 2003;9(9):1058-1063.
[3] Frank Esper, MD, Derek Boucher, et al. Human Metapneumovirus Infection in the United States: Clinical Manifestations Associated With a Newly Emerging Respiratory Infection in Children [J]. Pediatrics, 2003, 6(111) : 1407-1410.
[4] Nathalie Bastien, Diane Ward, Paul Van Caeseele, et al. Human Metapneumovirus Infection in the Canadian Population [J]. J. Clin. Microbiol., 2003; 10(41):4642- 4646.
[5] van den Hoogen, B. G., T. M. Bestebroer, et al. Analysis of the genomic sequence of a human metapneumovirus[J]. Virology, 2002; 295:119-132.
[6] Boivin G, Mackay I, Sloots TP, et al. Global genetic diversity of humanmetapneumovirus fusion gene [J]. Emerg Infect Dis, 2004; 10:1154–1157.
[7] Biacchesi S, Skiadopoulos MH, Boivin G, et al. Genetic diversity between human metapneumovirus subgroups[J]. Virology,2003; 315:1–9.
[8] Gregory CG, Ana WC, Sharon F, et al. Multi-year study of human metapneumovirus infection at a large US midwestern medical referral center [J]. J Clin Virol. 2006; 37(4):269-276.
[9] Murray CJL, Lopez AD, Mathers CD, et al. The global burden of disease 2000 project : aim, methods and data sources. Global programme on evidence for health policy.Geneva: World Health org, 2001,136.
[10] Ordas J, Boga JA, Alvarez-Arguelles M, et a1. Role of metapneumovlrus in viral respiratory infections in young children [J]. J Clin Microbiol 2006, 44(8): 2739-2742
[11] Mackay IM, Bialasiewicz S, Waliuzzaman Z, et a1. Use of the P gene to genotype human metapneumovirus identifies 4 viral subtypes [J] .J Infect Dis, 2004, 190 (11):1913-1918.
[12] Scheltinga SA, Templeton KE, Beemma MF, et a1. Diagnosis of human metapneumovirus and rhinovirus in patients with respiratory tract infections by an internally controlled multiplex real-time RNA PCR [J]. J Clin Virol, 2005, 33(4): 306-311.
[13] Sumino KC,Asapov E,Pierce RA , et a1.Detection of severe human metapneumovirus infection by real-time polymerase chain reaction and histopatho-logical assessment [J].J Infect Dis, 2005, 192(6):1052—1060.
[14]朱汝南,钱渊,赵林清,等.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.
[15]陈慧中,钱渊,王天有,等.偏肺病毒毛细支气管炎的临床研究[J].中华儿科杂志, 2004, 42(5):383-386.
[16] Peiris JS, TangWH, Chart KH, et al. Children with respiratory disease associated with metapneumovirus in HongKong[J].Emerg Infect Dis, 2003, 9(6): 628-633.
[17] van den Hoogen BG, Osterhaus DM, Fouchier RA. Clinical impact and diagnosis of human metapneumovirus infection[J].Pediatr Infect Dis J, 2004, 23:25-32
[18] van den Hoogen BG, van Doomum GJ, Fockem JC, et al. Prevalence and clinical symptoms of human metapneumovirus infecion in hospitalized patients[J]. J Infect Dis, 2003, 188:1571-1577.
[19] Esper F, Martinello RA, Boucher D, et al. A 1-year experience with human metapneumovirus in children aged<5 years[J]. J Infect Dis, 2004, 189: 1388-1396.
[20] Bastien N, Ward D, Van Caeseele PK, et al. Human metapneumovirus infection in the canadian population [J].J Clin Microbiol, 2003, 41(10): 4642-4646.
[21] Peiris JS, TangWH, Chart KH, et al. Children with respiratory disease associated with metapneumovirus in HongKong [J].Emerg Infect Dis, 2003, 9(6):628-633.
[22] Gerna G, Campanini G, Rovida F, et al. Changing circulation rate of human metapneumovirus stains and types among hospitalized pediatric patients during three consecutive winter-spring seasons[J]. Arch Virol, 2005, 150: 2365-2375.
[23] Greensill J,McNamara PS, Dove W, et al. Human metapneumovirus in sever respiratory syncytial virus bronchiolitis[J]. Emerg infect Dis 2003,9:372-375.
[24] kuypers J, Wright N, Corey L, et al. Detection and quantification of human metapneumovirus in pediatric specimens by real-time RT-PCR [J]. J Clin Virol 2005,33:299-305.
[25] Semple MG, Cowell A, Dove W, et al. Dual infection of infants by human metapneumovirus and human respiratory syncytial virus is strongly associated with severe bronchiolitis[J].J Infect Dis ,2005, 191:382-386.
[26] Nissen MD, Siebert DJ, Mackay IM, et al. Evidence of human metapneumovirus in Australian children[J]. Med J Aust 2002,176,:188.
[27] Boivin G, Abed Y, Pelletier G, et al.Virological features and clinicalmanifestations associated with human metapneumovirus:a new paramyxovirus responsible for acute respiratory-tract infections in all age groups[J].J.Infect.Dis. 2002, 186:1130-1334.
[1] van den Hoogen BG, deJong JC, Groen J, et al. A newly discovered human pneumovirus isolated from young children with respiratory tract disease [J]. Nat Med, 2001; 7: 719–724.
[2] Chan PKS, Tam JS, Lam C-W, et al. Human metapneumovirus detection in patients with severe acute respiratory syndrome [J]. Emerg Infect Dis, 2003;9(9):1058-1063.
[3] Frank Esper, MD, Derek Boucher, et al. Human Metapneumovirus Infection in the United States: Clinical Manifestations Associated With a Newly Emerging Respiratory Infection in Children [J]. Pediatrics,2003;6(111) : 1407-1410.
[4] Nathalie Bastien, Diane Ward, Paul Van Caeseele, et al. Human Metapneumovirus Infection in the Canadian Population [J]. J. Clin. Microbiol., 2003; 10(41):4642-4646.
[5] Biacchesi S, Skiadopoulos MH, Boivin G, et a1. Genetic diversity between human pneumovirus subground[J]. Virology. 2003, 315:1-9.
[6] van den Hoogen, B. G., T. M. Bestebroer, et al. Analysis of the genomic sequence of a human metapneumovirus[J]. Virology, 2002; 295:119-132.
[7] Boivin G, Abed Y, Pelletier G, et al.Virological features and clinical manifestations associated with human metapneumovirus:a new paramyxovirus responsible for acute respiratory-tract infections in all age group[J]s.J.Infect.DIS .2002, 186:1130-1334.
[8] Pelletier G, Dery P, Abed Y, Boivin G.Respiratory tract reinfections by the new human metapneumovirus in an immunocompromised child. Emerg . Infect . Dis . 2002, 8:976-978.
[9] Williams J, Harris P, Tollefson S, et al. Human metapneumovirus and lower respiratory tract disease in otherwise healthy infants and children, N. Engl . J . Med. 2004,350:443-450.
[10] van den Hoogen BG, Osterhaus DM, Fouchier RA. Clinical impact and diagnosis of human metapneumovirus infection[J].Pediatr Infect Dis J, 2004, 23: 25-32.
[11] van den Hoogen BG, van Doomum GJ, Fockem JC, et al. Prevalence and clinical symptoms of human metapneumovirus infecion in hospitalized patients[J]. J Infect Dis, 2003, 188:1571-1577.
[12] Esper F, Martinello RA, Boucher D, et al. A 1-year experience with human metapneumovirus in children aged<5 years[J]. J Infect Dis, 2004, 189: 1388-1396.
[13] Bastien N, Ward D, Van Caeseele PK, et al. Human metapneumovirus infection in the canadian population [J]. J Clin Microbiol, 2003, 41(10): 4642-4646.
[14] Peiris JS, TangWH, Chart KH, et al. Children with respiratory disease associated with metapneumovirus in HongKong[J].Emerg Infect Dis, 2003, 9(6):628-633.
[15]朱汝南,钱渊,邓洁,等.北京地区六岁以下儿童急性呼吸道偏肺病毒感染.中华儿科杂志[J],2003,41:441-444.
[16] Dollner H, Risnes K, Radtke A, et al. Outbreak of human metapneumovirus infection in Norwegian children[J]. Pediatr, J Infect Dis, 2004, 23:436-439.
[17] Mackay IM,Bialasiewicz S,Waliuzzaman Z, et a1.Use of the P gene to genotype human metapneumovirus identifies 4 viral subtypes [J] .J Infect Dis, 2004, 190 (11):1913-1918.
[18] Scheltinga SA, Templeton KE, Beemma MF, et a1.Diagnosis of human metapneumovirus and rhinovirus in patients with respiratory tract infections by an internally controlled multiplex real-time RNA PCR [J]. J Clin Virol, 2005, 33(4): 306-311.
[19] Sumino KC,Asapov E,Pierce RA , et a1.Detection of severe human metapneumovirus infection by real-time polymerase chain reaction and histopatho-logical assessment [J].J Infect Dis, 2005, 192(6):1052—1060.
[20] Louis JK,Hacker JK,Gonzales R,et a1.Characterization of viral agents causing acute respiratory infection in a San Francisco University Medical Center Clinic during the influenza season[J].Clin Infect Dis,2005,41(6):822-828.
[21] Noyola DE,Alpuche-Solis AG,Herrera-Diaz A,et a1.Human metapneum- ovirus infections in Mexico: epidemiological and clinical characteristics [J]. J Med Microbiol, 2005,54:969-974.
[22]陈慧中,钱渊,王天有,等.偏肺病毒毛细支气管炎的临床研究[J].中华儿科杂志,2004,42(5):383—386.
[23] Esper F, Boucher D, Weibel C, et a1. Human metapneumovirus infection in the United States:clinical manifestation associated with a newly emerging respiratory infection in children[J]. Pediatrics, 2003, 111:1407-1410.
[24] Rawlinson W,Waliuzzaman Z,Carter I ,et al.Asthma exacerbations in children associated with rhinovirus but no human metapneumovirus infection[J] J.Infect. Dis.2003,187:1314-1318.
[25] Williams J, Harris P, Tollefson S, et al. Human metapneumovirus and lower respiratory tract disease in otherwise healthy infants and children, N. Engl. J Med.2004,350:443-450.
[26] Carla P, van den Hoogen B, Chakraharti S, et al. Human metapneumovirus in a haematopoietic stem cell transplant recipient with fatal lower respiratory tractdiease[J]. Bone Marrow Transplant. 2003,31:309-310.
[27] Jartti T, Van den Hoogen B, Carofalo RP,et a1. Metapneumovirus and acute wheezing in children[J]. Lancet, 2002, 360(9343):1393-1394.
[28] Greensill J, McNamara P, Dove W , et al. Human metapneumovirus in severe respiratory syncytial virus bronchiolitis[J]. Emerg. Infect.Dis.2003,9 :372—375.
[29] Bosis S, Esposito S, Niesters H, et al. Impact of human metapneumovirus in childhood:Comparison with respiratory syncytial virus and influenza viruses[J]. J.Med virol .2005,75:101-104.
[30] Martinez I, Melero J. Binding of human respiratory syncytial virus to cells: implication of sulfated cell surface proteoglycans[J].J. Gen. Viro1.2000. 81: 2715-2722.
[31] Tripp R, Jones L, Haynes L, et al. CX3C chemokine mimicry by respiratory syncytial virus G glycoprotein. Nat. Immunol . 2001, 2:732-738.
[32] Peret T, Abed Y, Erdman D, et al. Sequence polymorphism of the predicted human metapneumovirus G glycoprotein [J].J. Gan. Viro1. 2004. 85:679-686.
[33] Alvarez R, Harrod K, Shieh W, et al. Human metapneumovirus persists in BALB/c mice despite the presence of neutralizing antibodies[J]. J.Virol. 2004. 78:14003-14011.