2016年～2017年山东地区商品肉鸡H9N2亚型禽流感病毒HA基因的分子特征和遗传进化分析
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摘要
为了解山东地区商品肉鸡H9N2亚型禽流感病毒(AIV)分离株的遗传变异情况,本研究对13株从不同肉鸡场分离的H9N2 AIV的HA基因进行PCR扩增、克隆和测序,并对获得的HA基因序列进行同源性和遗传进化分析。结果显示:13个分离株的HA裂解位点均为RSSR↓GLF,符合低致病性AIV的分子特征;8株分离株有8处糖基化位点,其余5株存在个别位点缺失情况;受体结合位点除198位有变异外,其它位点均保守;234位氨基酸均为L,因此具有与哺乳动物唾液酸α,2-6受体结合的特征。所有分离株HA基因核苷酸同源性为94.1%~99.8%,氨基酸同源性为94.3%~100%。HA基因进化树显示,所有分离株均属于欧亚分支中的A/Chicken/Beijing/1/1994(A/Duck/Hong Kong/Y280/97)亚群,也同属于近几年在中国鸡群中流行的以A/chicken/Zhejiang/HJ/2007为代表的G57分支。以上研究为H9N2亚型AIV的防控和疫苗研制提供了科学参考。
To understand the genetic variation of H9N2 subtype avian influenza virus(AIV) in Shandong province, HA genes of 13 isolates from different broiler farms were amplified, cloned and sequenced. The homology and genetic evolution of these HA genes were analyzed and showed that these isolates had a common motif RSSR ↓ GLF at the cleavage site which indicated that they were lentogenic. The deletion of glycosylation sites existed in five isolates, the other had eight glycosylation sites. In addition to the 198 th site, other receptor binding sites were conservative. The 234 amino acids were L, which had the characteristics of binding to mammalian sialic acid α,2-6 receptor. The nucleotide homologies of HA genes of the 13 isolates ranged from 94.1% to99.8%, and amino acid homologies were from 94.3% to 100%. Phylogenetic tree of HA genes analysis showed that all isolates belonged to the Eurasian branch of the A/Chicken/Beijing/1/1994(A/Duck/Hong Kong/Y280/97) subgroup, also belonged to the branch of G57, which have been prevalence in China among chick flocks in recent years represented by A/chicken/Zhejiang/HJ/2007. These data provided a reference for the prevention and control of H9N2 subtype AIVs and the vaccine development.
To understand the genetic variation of H9N2 subtype avian influenza virus(AIV) in Shandong province, HA genes of 13 isolates from different broiler farms were amplified, cloned and sequenced. The homology and genetic evolution of these HA genes were analyzed and showed that these isolates had a common motif RSSR ↓ GLF at the cleavage site which indicated that they were lentogenic. The deletion of glycosylation sites existed in five isolates, the other had eight glycosylation sites. In addition to the 198 th site, other receptor binding sites were conservative. The 234 amino acids were L, which had the characteristics of binding to mammalian sialic acid α,2-6 receptor. The nucleotide homologies of HA genes of the 13 isolates ranged from 94.1% to99.8%, and amino acid homologies were from 94.3% to 100%. Phylogenetic tree of HA genes analysis showed that all isolates belonged to the Eurasian branch of the A/Chicken/Beijing/1/1994(A/Duck/Hong Kong/Y280/97) subgroup, also belonged to the branch of G57, which have been prevalence in China among chick flocks in recent years represented by A/chicken/Zhejiang/HJ/2007. These data provided a reference for the prevention and control of H9N2 subtype AIVs and the vaccine development.
引文
[1]陈伯伦,张泽纪,陈伟斌.禽流感研究:鸡A型禽流感病毒的分离与血清学初步鉴定[J].中国兽医杂志,1994,22(10):3-5.
[2]Watanabe T,Kiso M,Fukuyama S,et al.Characterization of H7N9 influenza A viruses isolated from humans[J].Nature,2013,501(7468):551-555.
[3]Chen Hai-ying,Yuan Hui,Gao Rong-bao,et al.Clinical and epidemiological characteristics of a fatal case of avian influenza A H10N8 virus infection:a descriptive study[J].Lancet,2014,383(9918):714-721.
[4]万春和,傅光华,程龙飞,等.A/Chicken/Zhejiang/HJ/2007(H9N2)禽流感病毒8基因测序及遗传进化分析[J].农业生物技术学报,2009,17(5):750-757.
[5]石火英,孙蕾,陈素娟,等.H9N2亚型禽流感病毒复制特性的基因分析[J].畜牧兽医学报,2008,39(2):189-194.
[6]Rohm C,Horimoto T.Do hemagglutinin genes of highly pathogenic avian influenza virus constitute unique phylogenetic lineages?[J].Virology,1995,209(2):664-670.
[7]刘金华,吴清民,陈福勇,等.鸡源株与人源株H9N2流感病毒血凝素受体结合位点氨基酸比较与分析[J].中国预防兽医学报,2003,25(6):416-418.
[2]Watanabe T,Kiso M,Fukuyama S,et al.Characterization of H7N9 influenza A viruses isolated from humans[J].Nature,2013,501(7468):551-555.
[3]Chen Hai-ying,Yuan Hui,Gao Rong-bao,et al.Clinical and epidemiological characteristics of a fatal case of avian influenza A H10N8 virus infection:a descriptive study[J].Lancet,2014,383(9918):714-721.
[4]万春和,傅光华,程龙飞,等.A/Chicken/Zhejiang/HJ/2007(H9N2)禽流感病毒8基因测序及遗传进化分析[J].农业生物技术学报,2009,17(5):750-757.
[5]石火英,孙蕾,陈素娟,等.H9N2亚型禽流感病毒复制特性的基因分析[J].畜牧兽医学报,2008,39(2):189-194.
[6]Rohm C,Horimoto T.Do hemagglutinin genes of highly pathogenic avian influenza virus constitute unique phylogenetic lineages?[J].Virology,1995,209(2):664-670.
[7]刘金华,吴清民,陈福勇,等.鸡源株与人源株H9N2流感病毒血凝素受体结合位点氨基酸比较与分析[J].中国预防兽医学报,2003,25(6):416-418.