1株重组的H5N2亚型禽流感病毒全基因组序列分析
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摘要
本研究对1株H5N2亚型禽流感病毒(CK/GD02/14)进行全基因测序及遗传进化分析,结果显示,该病毒HA蛋白裂解位点含有多个连续的碱性氨基酸(321PQIEGRRRKR*GLF333),为高致病性禽流感病毒特征。遗传进化分析结果显示,CK/GD02/14与A/chicken/Jiangsu/1001/2013(H5N2)各基因片段都有较高的同源性(97.6%~98.9%)。HA基因位于H5N1亚型遗传进化树的7.2分支,NA基因属于H9N2亚型遗传进化树的BJ/1/94-like分支。其内部基因与1株H9N2亚型病毒的内部基因有较高的同源性,但其M基因属于类H5N1病毒分支,表明该H5N2亚型禽流感病毒为H9N2和H5N1亚型禽流感病毒的重组毒株。结果表明,CK/GD02/14与A/chicken/Jiangsu/1001/2013(H5N2)为同源毒株,可能由江苏传到广东地区。
The complete genome of CK/GD02/14 virus was amplified to sequence and the phylogenetic and molecular characteristics of the virus were analyzed.The results showed that the HA cleavage sequence of the virus possessed the 321PQIEGRRRKR*GLF333 motif,which indicated that the virus was highly pathogenic for chicken.Phylogenetic analysis showed that all the eight segment of CK/GD02/14 shared high homologies(97.6%-98.9%)with A/chicken/Jiangsu/1001/2013(H5N2).CK/GD02/14 possessed hemagglutinin(HA)gene belonged to clade 7.2H5 virus and neuraminidase(NA)gene belonged to BJ/1/94-like virus.Moreover,the internal genes of CK/GD02/14 were from H9N2 epidemic viruses,but the M gene belong to H5N1-like,which suggested that H9N2 viruses were generated by reassortance of H9N2 and H5N1viruses.These results indicated that CK/GD02/14 was homologous with CK/JS/1001/13,which suggested that the virus transmitted from Jiangsu province to Guangdong province.
The complete genome of CK/GD02/14 virus was amplified to sequence and the phylogenetic and molecular characteristics of the virus were analyzed.The results showed that the HA cleavage sequence of the virus possessed the 321PQIEGRRRKR*GLF333 motif,which indicated that the virus was highly pathogenic for chicken.Phylogenetic analysis showed that all the eight segment of CK/GD02/14 shared high homologies(97.6%-98.9%)with A/chicken/Jiangsu/1001/2013(H5N2).CK/GD02/14 possessed hemagglutinin(HA)gene belonged to clade 7.2H5 virus and neuraminidase(NA)gene belonged to BJ/1/94-like virus.Moreover,the internal genes of CK/GD02/14 were from H9N2 epidemic viruses,but the M gene belong to H5N1-like,which suggested that H9N2 viruses were generated by reassortance of H9N2 and H5N1viruses.These results indicated that CK/GD02/14 was homologous with CK/JS/1001/13,which suggested that the virus transmitted from Jiangsu province to Guangdong province.
引文
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[4]Stevens J,Blixt O,Tumpey T M,et al.Structure and receptor specificity of the hemagglutinin from an H5N1influenza virus[J].Science,2006,312(5772):404-410.
[5]Sun Y,Tan Y,Wei K,et al.Amino acid 316of hemagglutinin and the neuraminidase stalk length influence virulence of H9N2influenza virus in chickens and mice[J].J Virol,2013,87(5):2963-2968.
[6]Yao Y,Mingay L J,Mccauley J W,et al.Sequences in influenza A virus PB2protein that determine productive infection for an avian influenza virus in mouse and human cell lines[J].J Virol,2001,75(11):5410-5415.
[7]Li Z,Chen H,Jiao P,et al.Molecular basis of replication of duck H5N1influenza viruses in a mammalian mouse model[J].J Virol,2005,79(18):12058-12064.
[8]陈化兰,于康震.一株鹅源高致病力禽流感病毒分离株血凝素基因的分析[J].中国农业科学,1999,32(2):87-92.
[9]Gu M,Zhao G,Zhao K,et al.Novel variants of clade 2.3.4highly pathogenic avian influenza A(H5N1)viruses,China[J].Emerg Infect Dis,2013,19(12):2021-2024.
[10]Cheng M C,Soda K,Lee M S,et al.Isolation and characterization of potentially pathogenic H5N2influenza virus from a chicken in Taiwan in 2008[J].Avian Dis,2010,54(2):885-893.
[11]Lee J H,Pascua P N,Song M S,et al.Isolation and genetic characterization of H5N2influenza viruses from pigs in Korea[J].J Virol,2009,83(9):4205-4215.
[12]Zhan G J,Ling Z S,Zhu Y L,et al.Genetic characterization of a novel influenza A virus H5N2isolated from a dog in China[J].Vet Microbiol,2012,155(2/4):409-416.
[13]Ogata T,Yamazaki Y,Okabe N,et al.Human H5N2avian influenza infection in Japan and the factors associated with high H5N2-neutralizing antibody titer[J].J Epidemiol,2008,18(4):160-166.
[14]刘志刚,赵苏红,索朗斯珠,等.禽流感病毒H5N2亚型西藏株的分离鉴定与特性分析[J].中国兽医学报,2014,34(1):17-20.
[15]李广伟,严专强,廖昌韬,等.两广地区2011-2012年H9N2亚型禽流感病毒的HA基因进化分析[J].中国兽医学报,2014,34(3):461-464.
[16]Zhao K,Gu M,Zhong L,et al.Characterization of three H5N5and one H5N8highly pathogenic avian influenza viruses in China[J].Vet Microbiol,2013,163(3/4):351-357.
[17]Mi Z,Liu W,Fan H,et al.Complete Genome Sequence of Avian Influenza Virus A/chicken/Jiangsu/1001/2013(H5N2),Demonstrating Continuous Reassortance of H5N2in China[J].Genome Announc,2013,1(4):e00469-e00513.