类禽H1N1亚型与Pdm/09H1N1亚型猪流感重组病毒的分离鉴定及其遗传进化分析
|
摘要
为了解猪流感病毒(SIV)的流行情况,本试验在2014年10月-2015年9月期间在广东省各地区采集猪的鼻拭子和肺脏病料,将样品处理之后进行鸡胚分离和RT-PCR检测,分离到1株SIV,对其进行全基因组测序和遗传进化分析,结果表明为H1N1亚型SIV,病毒HA,NA,M和NS基因片段属于类禽分支(EA),病毒PB2、PB1、PA和NP基因片段属于Pdm/09分支。HA基因序列同源性结果显示,核苷酸相似性为93.2%~99.1%,其中与A/swine/Jiangxi/3858/2011(H1N1)SIV同源性最高。抗原位点分析结果表明,分离毒株的抗原位点基本保守。本研究通对广东省各地区分离的SIV分析,为SIV的重组和变异趋势及公共卫生安全的防控提供依据。
To investigate the epidemiclogy and variation of Swine influenza virus(SIV)in Guangdong province,a total of 1 259 specimens included nasal swab and lung issue were collected from pigs in 4 areas of Guangdong province from Nov/2014 to Sept/2015.After processed specimens,SIV virus strain was isolated by inoculation in 10-days-old embryonated eggs and RT-PCR test.The complete genes were sequenced and the genetic analysis showed that the virus strain was H1N1 SIV virus.The genetic analysis showed it was close related to A/Jiangxi/3858/2011(H1N1),the nucleotide identity was between 93.2%-99.1%.The phylogenetic analysis showed that PB2,PB1,PA and NP of virus strains belong to Pdm/09 branch,The external gene HA and NA,NS gene and M gene belong to Europe-Asia-avian-like strain.The result of aantigenic site analysis demonstrated that antigenic site of the isolate was generally conserved.This study made an effort to survey the mutation trend of SIV and provided laboratory data for prevention of SIV.
To investigate the epidemiclogy and variation of Swine influenza virus(SIV)in Guangdong province,a total of 1 259 specimens included nasal swab and lung issue were collected from pigs in 4 areas of Guangdong province from Nov/2014 to Sept/2015.After processed specimens,SIV virus strain was isolated by inoculation in 10-days-old embryonated eggs and RT-PCR test.The complete genes were sequenced and the genetic analysis showed that the virus strain was H1N1 SIV virus.The genetic analysis showed it was close related to A/Jiangxi/3858/2011(H1N1),the nucleotide identity was between 93.2%-99.1%.The phylogenetic analysis showed that PB2,PB1,PA and NP of virus strains belong to Pdm/09 branch,The external gene HA and NA,NS gene and M gene belong to Europe-Asia-avian-like strain.The result of aantigenic site analysis demonstrated that antigenic site of the isolate was generally conserved.This study made an effort to survey the mutation trend of SIV and provided laboratory data for prevention of SIV.
引文
[1]YANG H,CHEN Y,QIAO C,et al.Prevalence,genetics,and transmissibility in ferrets of Eurasian avianlike H1N1swine influenza viruses[J].PNAS,2015,113(2):392.
[2]唐雨博,徐文章,孙艺学,等.H9N2亚型流感病毒血凝素受体结合区定点突变株拯救及其受体亲和性鉴定[J].中国兽医学报,2017,37(7):1243-1248.
[3]金新,于洪敏,张茂林,等.禽流感病毒AM2蛋白相互作用宿主蛋白的筛选及初步验证[J].中国兽医学报,2017,37(5):866-870.
[4]YAN C,JIAN Z,QIAO C L,et al.Co-circulation of pandemic 2009H1N1,classical swine H1N1and avian-like swine H1N1influenza viruses in pigs in China[J].Infect Genet Evo,2013,13(23):331-338.
[5]SONG M S,HAN L J,PASCUA P N Q,et al.Evidence of human-to-swine transmission of the pandemic(h1n1)2009influenza virus in south korea[J].J Clin Microbiol,2010,48(9):3204-3211.
[6]FAN X,ZHU H,ZHOU B,et al.Emergence and dissemination of a swine H3N2reassortant influenza virus with 2009pandemic H1N1genes in pigs in china[J].J Virol,2012,86(4):2375-2378.
[7]CHEN J,MA J,WHITE S K,et al.Live poultry market workers are susceptible to both avian and swine influenza viruses,Guangdong Province,China[J].Vet Microbiol,2015,181(3/4):230-235.
[8]LIU W,WEI M T,TONG Y,et al.Seroprevalence and genetic characteristics of five subtypes of influenza A viruses in the Chinese pig population:A pooled data analysis[J].Vet J,2011,187(2):200-206.
[9]DUCATEZ M F,HAUSE B,STIGGERROSSER E,et al.Multiple reassortment between pandemic(H1N1)2009and endemic influenza viruses in pigs,united states[J].Emerg Infect Dis,2011,17(9):1624-1629.
[10]MORENO A,DI T L,FACCINI S,et al.Novel H1N2swine influenza reassortant strain in pigs derived from the pandemic H1N1/2009virus[J].Vet Microbiol,2011,149(3/4):472-477.
[11]ZHU H,ZHOU B,FAN X,et al.Novel reassortment of eurasian avian-like and pandemic/2009influenza viruses in swine:infectious potential for humans[J].J Virol,2011,85(20):10432-10439.
[12]QIAO C,LIU L,YANG H,et al.Novel triple reassortant H1N2influenza viruses bearing six internal genes of the pandemic 2009/H1N1influenza virus were detected in pigs in China[J].J Clin Virol,2014,61(4):529-534.
[13]HOFFMAN L M.The return of the city-state:urban governance and the Yew York City H1N1Pandemic[J].Sociol Health Illn,2013,35(2):255-267.
[14]李爽,王小攀.一类变异为H7N9型禽流感病毒模型的稳定性分析[J].扬州大学学报(自然科学版),2016(4):14-17.
[15]戴华,徐正中,陈俊华,等.H5N1亚型禽流感灭活疫苗和DNA疫苗诱导IFN-γ应答评价[J].扬州大学学报(农业与生命科学版),2017,38(3):1-5.
[16]肖越胜,宿春虎,黄瑶,等.扬州市近3年新甲型H1N1流感病毒HA基因分析[J].扬州大学学报(农业与生命科学版),2013,34(1):36-40.
[17]SUI J,YANG D,QIAO C,et al.Protective efficacy of an inactivated Eurasian avian-like H1N1swine influenza vaccine against homologous H1N1and heterologous H1N1and H1N2viruses in mice[J].Vaccine,2016,34(33):3757-3763.
[18]GARTEN R J,DAVIS C T,RUSSELL,C A et al.Antigenic and genetic characteristics of swine-origin 2009a(H1N1)influenza viruses circulating in humans[J].Science,2009,5937(325):197-201.
[19]彭特,许利娜,张思远,等.1株HA蛋白S145变异H9N2亚型禽流感病毒的分离鉴定与HA基因序列分析[J].中国兽医学报,2017,37(4):676-681.
[2]唐雨博,徐文章,孙艺学,等.H9N2亚型流感病毒血凝素受体结合区定点突变株拯救及其受体亲和性鉴定[J].中国兽医学报,2017,37(7):1243-1248.
[3]金新,于洪敏,张茂林,等.禽流感病毒AM2蛋白相互作用宿主蛋白的筛选及初步验证[J].中国兽医学报,2017,37(5):866-870.
[4]YAN C,JIAN Z,QIAO C L,et al.Co-circulation of pandemic 2009H1N1,classical swine H1N1and avian-like swine H1N1influenza viruses in pigs in China[J].Infect Genet Evo,2013,13(23):331-338.
[5]SONG M S,HAN L J,PASCUA P N Q,et al.Evidence of human-to-swine transmission of the pandemic(h1n1)2009influenza virus in south korea[J].J Clin Microbiol,2010,48(9):3204-3211.
[6]FAN X,ZHU H,ZHOU B,et al.Emergence and dissemination of a swine H3N2reassortant influenza virus with 2009pandemic H1N1genes in pigs in china[J].J Virol,2012,86(4):2375-2378.
[7]CHEN J,MA J,WHITE S K,et al.Live poultry market workers are susceptible to both avian and swine influenza viruses,Guangdong Province,China[J].Vet Microbiol,2015,181(3/4):230-235.
[8]LIU W,WEI M T,TONG Y,et al.Seroprevalence and genetic characteristics of five subtypes of influenza A viruses in the Chinese pig population:A pooled data analysis[J].Vet J,2011,187(2):200-206.
[9]DUCATEZ M F,HAUSE B,STIGGERROSSER E,et al.Multiple reassortment between pandemic(H1N1)2009and endemic influenza viruses in pigs,united states[J].Emerg Infect Dis,2011,17(9):1624-1629.
[10]MORENO A,DI T L,FACCINI S,et al.Novel H1N2swine influenza reassortant strain in pigs derived from the pandemic H1N1/2009virus[J].Vet Microbiol,2011,149(3/4):472-477.
[11]ZHU H,ZHOU B,FAN X,et al.Novel reassortment of eurasian avian-like and pandemic/2009influenza viruses in swine:infectious potential for humans[J].J Virol,2011,85(20):10432-10439.
[12]QIAO C,LIU L,YANG H,et al.Novel triple reassortant H1N2influenza viruses bearing six internal genes of the pandemic 2009/H1N1influenza virus were detected in pigs in China[J].J Clin Virol,2014,61(4):529-534.
[13]HOFFMAN L M.The return of the city-state:urban governance and the Yew York City H1N1Pandemic[J].Sociol Health Illn,2013,35(2):255-267.
[14]李爽,王小攀.一类变异为H7N9型禽流感病毒模型的稳定性分析[J].扬州大学学报(自然科学版),2016(4):14-17.
[15]戴华,徐正中,陈俊华,等.H5N1亚型禽流感灭活疫苗和DNA疫苗诱导IFN-γ应答评价[J].扬州大学学报(农业与生命科学版),2017,38(3):1-5.
[16]肖越胜,宿春虎,黄瑶,等.扬州市近3年新甲型H1N1流感病毒HA基因分析[J].扬州大学学报(农业与生命科学版),2013,34(1):36-40.
[17]SUI J,YANG D,QIAO C,et al.Protective efficacy of an inactivated Eurasian avian-like H1N1swine influenza vaccine against homologous H1N1and heterologous H1N1and H1N2viruses in mice[J].Vaccine,2016,34(33):3757-3763.
[18]GARTEN R J,DAVIS C T,RUSSELL,C A et al.Antigenic and genetic characteristics of swine-origin 2009a(H1N1)influenza viruses circulating in humans[J].Science,2009,5937(325):197-201.
[19]彭特,许利娜,张思远,等.1株HA蛋白S145变异H9N2亚型禽流感病毒的分离鉴定与HA基因序列分析[J].中国兽医学报,2017,37(4):676-681.