2017年冬季上海地区犬流感病毒的检测与HA、NA序列的遗传进化分析
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摘要
为了解2017年冬季上海临床接诊犬的流感病毒感染情况和病毒变异趋势,收集上海某动物医院疑似犬流感样品进行RT-PCR检测,构建HA、NA的重组载体并测序,对获得的序列进行进化分析、氨基酸变异分析,综合我国同亚型犬流感病毒HA序列进行进化率分析。结果显示:23份疑似样品中检出5份阳性样品,获得5个H3亚型的HA序列、2个N2亚型的NA序列,上述序列在进化树中已经形成独立的进化分支,与韩国、广东毒株亲缘关系最近;氨基酸比对表明,HA和NA在重要位点发生许多一致的突变,如抗原位点区域、糖基化位点等,NA茎部未见氨基酸插入;对2007年至2017年中国南方地区分离的H3N2犬流感病毒HA片段进行进化率、变异率分析显示,年平均进化率呈逐年增高趋势,变异不断积累。研究表明,开展犬流感病毒变异监测,掌握病毒变异和进化方向,对防控该病、防止病毒向人群传播有重要意义。
Canine influenza virus( CIV) is a newly emerged infectious disease. However,little is known about the prevalence and variation of this virus in pet dogs,and this study was aimed at the surveillance of CIV infection in a veterinary clinic of Shanghai. Five positive samples were obtained by RT-PCR from 23 nasopharyngeal swabs. Five HA segments of H3 subtype and 2 NA segments of N2 subtype were sequenced using a recombinant T-vector. From the phylogenetic trees,the H3 segments identified in this study were most similar to H3N2 viruses isolated in Korea and Guangdong Province and were clustered into an independent clade,and so were the N2 segments. Comparison of the deduced amino acid sequences showed that all the segments had several mutations at important positions. In HA1,there were some substitutions at the antigenic sites,glycosylation sites and cleavage sites. As for NA,no insertion in stalk was found. Furthermore,calculation with HA sequences isolated in southern China from 2007 to 2017 revealed that the average evolution rate increased in the line chart indicating accumulation of antigen drift. Overall,these data indicated that in-depth study was required for a better understanding of the variation of CIV and control of its potential threat to public health.
Canine influenza virus( CIV) is a newly emerged infectious disease. However,little is known about the prevalence and variation of this virus in pet dogs,and this study was aimed at the surveillance of CIV infection in a veterinary clinic of Shanghai. Five positive samples were obtained by RT-PCR from 23 nasopharyngeal swabs. Five HA segments of H3 subtype and 2 NA segments of N2 subtype were sequenced using a recombinant T-vector. From the phylogenetic trees,the H3 segments identified in this study were most similar to H3N2 viruses isolated in Korea and Guangdong Province and were clustered into an independent clade,and so were the N2 segments. Comparison of the deduced amino acid sequences showed that all the segments had several mutations at important positions. In HA1,there were some substitutions at the antigenic sites,glycosylation sites and cleavage sites. As for NA,no insertion in stalk was found. Furthermore,calculation with HA sequences isolated in southern China from 2007 to 2017 revealed that the average evolution rate increased in the line chart indicating accumulation of antigen drift. Overall,these data indicated that in-depth study was required for a better understanding of the variation of CIV and control of its potential threat to public health.
引文
[1] Biggerstaff M,Cauchemez S,Reed C,et al. Estimates of the reproduction number for seasonal,pandemic,and zoonotic influenza:a systematic review of the literature[J]. Bmc Infect Dis,2014,14(1):480.
[2] Harder T C,Vahlenkamp T W. Influenza virus infections in dogs and cats[J]. Vet Immunol Immunop,2010,134(1/2):54-60.
[3]夏冰,刘红彦,胡国良,等.犬流感的研究进展及其公共卫生安全影响[J].中国动物检疫,2014(10):33-37.
[4] Crawford P C,Dubovi E J,Castleman W L,et al. Transmission of equine influenza virus to dogs[J]. Sci, 2005, 310(5747):482-485.
[5] Nakajima K,Nobusawa E,Tonegawa K,et al. Restriction of amino acid change in influenza A virus H3HA:comparison of amino acid changes observed in nature and in vitro[J]. J Virol,2003,77(18):10088-10098.
[6] Su S,Chen Y,Zhao F R,et al. Avian-origin H3N2 canine influenza virus circulating in farmed dogs in Guangdong,China[J].Infect Genet Evol,2013,14(2):444-449.
[7] Song D,Kang B,Lee C,et al. Transmission of avian influenza virus(H3N2)to dogs[J]. Emerg Infect Dis,2008,14(5):741-746.
[8] Huang Q,Sivaramakrishna R P,Ludwig K,et al. Early steps of the conformational change of influenza virus hemagglutinin to a fusion active state:stability and energetics of the hemagglutinin[J]. BBABiophys,2003,1614(1):3-13.
[9] Palese P,Tobita K,Ueda M,et al. Characterization of temperature sensitive influenza virus mutants defective in neuraminidase[J].Virology,1974,61(2):397-410.
[10]杜念兴,潘芹芹,张素芳.遗传距离的错误观点和病毒进化踪迹的探索[J].中国病毒学,2005(3):335-339.
[11]闫中山,洪马林,付橙等.一株H3N2亚型犬流感病毒的分离鉴定及遗传进化分析[J].中国预防兽医学报,2016(5):407-409.
[12] Xu R,Ekiert D C,Krause J C,et al. Structural basis of preexisting immunity to the 2009 H1N1 pandemic influenza virus[J]. Sci,2010,328(5976):357-360.
[13] Kuiken T,Holmes E C,Mc Cauley J,et al. Host species barriers to influenza virus infections[J]. Sci, 2006, 312(5772):394-397.
[14]孟芳,徐怀英,张伟等.近20年中国部分地区鸡源H9N2亚型禽流感病毒HA基因遗传演化及其变异频率[J].微生物学报,2016(1):35-43.
[15] Pan C,Wang G,Liao M,et al. High genetic and antigenic similarity between a swine H3N2 influenza A virus and a prior human influenza vaccine virus:a possible immune pressure-driven cross-species transmission[J]. Biochem Bioph Res Co,2009,385(3):402-407.
[16] Sun L,Zhang G,Shu Y,et al. Genetic correlation between H3N2human and swine influenza viruses[J]. J Clin Virol,2009,44(2):141-144.
[17] Chang C P,New A E,Taylor J F,et al. Influenza virus isolations from dogs during a human epidemic in Taiwan[J]. Inter J Zoonoses,1976,3(1):61-64.
[18] Ito T,Couceiro J N,Kelm S,et al. Molecular basis for the generation in pigs of influenza A viruses with pandemic potential[J]. J Virol,1998,72(9):7367-7373.
[19] Lees W D,Moss D S,Shepherd A J. A computational analysis of the antigenic properties of haemagglutinin in influenza A H3N2[J].Bioinformatics,2010,26(11):1403-1408.
[20] Ito T. Interspecies transmission and receptor recognition of influenza A viruses[J]. Microbiol Immunol,2000,44(6):423-430.
[21]林焱. H3N2亚型犬流感病毒江苏分离株基因组特征与致病特性研究[D].南京:南京农业大学,2014:53.
[22] Steinhauer D A. Role of hemagglutinin cleavage for the pathogenicity of influenza virus[J]. Virol,1999,258(1):1-20.
[23]黄维娟,成艳辉,李希妍,等. 2011-2012年度中国H3N2亚型流感病毒病原学特征分析[J].病毒学报,2013(3):258-264.
[24] Huang J W,King C C,Yang J M. Co-evolution positions and rules for antigenic variants of human influenza A/H3N2 viruses[J].BMC Bioinformatics,2009,10(Suppl 1):S41.
[25] Mesman A W,Westerhuis B M,Ten Hulscher H I,et al. Influenza virus A(H1N1)2009 antibody-dependent cellular cytotoxicity in young children prior to the H1N1 pandemic[J]. J Gen Virol,2016,97(9):2157-2165.
[26] Baigent S J,Mc Cauley J W. Glycosylation of haemagglutinin and stalk-length of neuraminidase combine to regulate the growth of avian influenza viruses in tissue culture[J]. Virus Res,2001,79(1/2):177-185.
[27] Castrucci M R,Kawaoka Y. Biologic importance of neuraminidase stalk length in influenza A virus[J]. J Virol,1993,67(2):759-764.
[28] Lin Y,Zhao Y,Zeng X,et al. Genetic and pathobiologic characterization of H3N2 canine influenza viruses isolated in the Jiangsu Province of China in 2009-2010[J]. Vet Microbiol,2012,158(3/4):247-258.
[2] Harder T C,Vahlenkamp T W. Influenza virus infections in dogs and cats[J]. Vet Immunol Immunop,2010,134(1/2):54-60.
[3]夏冰,刘红彦,胡国良,等.犬流感的研究进展及其公共卫生安全影响[J].中国动物检疫,2014(10):33-37.
[4] Crawford P C,Dubovi E J,Castleman W L,et al. Transmission of equine influenza virus to dogs[J]. Sci, 2005, 310(5747):482-485.
[5] Nakajima K,Nobusawa E,Tonegawa K,et al. Restriction of amino acid change in influenza A virus H3HA:comparison of amino acid changes observed in nature and in vitro[J]. J Virol,2003,77(18):10088-10098.
[6] Su S,Chen Y,Zhao F R,et al. Avian-origin H3N2 canine influenza virus circulating in farmed dogs in Guangdong,China[J].Infect Genet Evol,2013,14(2):444-449.
[7] Song D,Kang B,Lee C,et al. Transmission of avian influenza virus(H3N2)to dogs[J]. Emerg Infect Dis,2008,14(5):741-746.
[8] Huang Q,Sivaramakrishna R P,Ludwig K,et al. Early steps of the conformational change of influenza virus hemagglutinin to a fusion active state:stability and energetics of the hemagglutinin[J]. BBABiophys,2003,1614(1):3-13.
[9] Palese P,Tobita K,Ueda M,et al. Characterization of temperature sensitive influenza virus mutants defective in neuraminidase[J].Virology,1974,61(2):397-410.
[10]杜念兴,潘芹芹,张素芳.遗传距离的错误观点和病毒进化踪迹的探索[J].中国病毒学,2005(3):335-339.
[11]闫中山,洪马林,付橙等.一株H3N2亚型犬流感病毒的分离鉴定及遗传进化分析[J].中国预防兽医学报,2016(5):407-409.
[12] Xu R,Ekiert D C,Krause J C,et al. Structural basis of preexisting immunity to the 2009 H1N1 pandemic influenza virus[J]. Sci,2010,328(5976):357-360.
[13] Kuiken T,Holmes E C,Mc Cauley J,et al. Host species barriers to influenza virus infections[J]. Sci, 2006, 312(5772):394-397.
[14]孟芳,徐怀英,张伟等.近20年中国部分地区鸡源H9N2亚型禽流感病毒HA基因遗传演化及其变异频率[J].微生物学报,2016(1):35-43.
[15] Pan C,Wang G,Liao M,et al. High genetic and antigenic similarity between a swine H3N2 influenza A virus and a prior human influenza vaccine virus:a possible immune pressure-driven cross-species transmission[J]. Biochem Bioph Res Co,2009,385(3):402-407.
[16] Sun L,Zhang G,Shu Y,et al. Genetic correlation between H3N2human and swine influenza viruses[J]. J Clin Virol,2009,44(2):141-144.
[17] Chang C P,New A E,Taylor J F,et al. Influenza virus isolations from dogs during a human epidemic in Taiwan[J]. Inter J Zoonoses,1976,3(1):61-64.
[18] Ito T,Couceiro J N,Kelm S,et al. Molecular basis for the generation in pigs of influenza A viruses with pandemic potential[J]. J Virol,1998,72(9):7367-7373.
[19] Lees W D,Moss D S,Shepherd A J. A computational analysis of the antigenic properties of haemagglutinin in influenza A H3N2[J].Bioinformatics,2010,26(11):1403-1408.
[20] Ito T. Interspecies transmission and receptor recognition of influenza A viruses[J]. Microbiol Immunol,2000,44(6):423-430.
[21]林焱. H3N2亚型犬流感病毒江苏分离株基因组特征与致病特性研究[D].南京:南京农业大学,2014:53.
[22] Steinhauer D A. Role of hemagglutinin cleavage for the pathogenicity of influenza virus[J]. Virol,1999,258(1):1-20.
[23]黄维娟,成艳辉,李希妍,等. 2011-2012年度中国H3N2亚型流感病毒病原学特征分析[J].病毒学报,2013(3):258-264.
[24] Huang J W,King C C,Yang J M. Co-evolution positions and rules for antigenic variants of human influenza A/H3N2 viruses[J].BMC Bioinformatics,2009,10(Suppl 1):S41.
[25] Mesman A W,Westerhuis B M,Ten Hulscher H I,et al. Influenza virus A(H1N1)2009 antibody-dependent cellular cytotoxicity in young children prior to the H1N1 pandemic[J]. J Gen Virol,2016,97(9):2157-2165.
[26] Baigent S J,Mc Cauley J W. Glycosylation of haemagglutinin and stalk-length of neuraminidase combine to regulate the growth of avian influenza viruses in tissue culture[J]. Virus Res,2001,79(1/2):177-185.
[27] Castrucci M R,Kawaoka Y. Biologic importance of neuraminidase stalk length in influenza A virus[J]. J Virol,1993,67(2):759-764.
[28] Lin Y,Zhao Y,Zeng X,et al. Genetic and pathobiologic characterization of H3N2 canine influenza viruses isolated in the Jiangsu Province of China in 2009-2010[J]. Vet Microbiol,2012,158(3/4):247-258.
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