一株鸭源H10N5亚型禽流感病毒的全基因组序列分析及其对小鼠的感染性研究
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摘要
为了解湖北省活禽市场鸭源H10N5亚型禽流感病毒(AIV) A/duck/Hu B/S1254/2014 (H10N5)株的生物学特性,本研究对其全基因组进行了测序,并以BABL/c小鼠为动物模型评价其对哺乳动物的感染性。序列分析结果显示:该病毒HA蛋白的裂解位点为334PELMQGR↓GLF343,符合低致病性AIV的特征。其表面基因HA、NA以及内部基因PB2和PA与1株江西野鸟源病毒株A/migratory duck/Jiangxi/30246/2013 (H10N5)高度同源,但其它内部基因节段PB1、NP、M和NS则来源于多个亚型的AIV。对小鼠的感染性实验结果显示,该病毒不需要提前适应就能够在小鼠的鼻甲和肺脏中有效复制,病毒滴度分别为4.08 log10 EID50/m L,5.0 log10 EID50/mL,病毒对小鼠呈低致病性,小鼠体重无显著的下降趋势,在感染期间小鼠未出现明显临床症状。本研究为了解H10N5亚型AIV分子进化关系,评估其对哺乳动物的感染风险提供了数据支持。
H10 N5 avian influenza virus(AIV) of A/Duck/HuB/S1254/2014 was isolated from duck in a live poultry market in Hubei province during the routine epidemiologic surveillance in 2014. The virus complete genome was sequenced to know biological characteristics, the pathogenicity of the AIV was studied in BABL/c mice as a model, which was evaluated for mammalian infectivity. Sequence analysis of the hemagglutinin(HA) gene results showed that the virus was a characteristic of low pathogenic AIV, with the typical HA cleavage site of 334 PELMQGR↓GLF343. The highest nucleotide sequence of HA, NA, PA and PB2 genes were similar to A/migratoryduck/Jiangxi/30246/2013(H10 N5). However, the other internal gene segments of PB1, NP,M and NS genes originated from multiple subtypes. Pathogenicity experiments showed that the virus could be replicated in the turbinate and lung of mice without prior adaptation, with virus titers of 4.08 log10 EID50/mL and 5.0 log10 EID50/mL, respectively,The virus showed low pathogenicity to the mice, no obvious clinical symptoms and significant change of body weight during infection in mice. The results will contribute to understand the molecular evolutionary relationship of H10 N5 AIV and provide areference for evaluating the potential threat of H10 N5 AIV infection in mammals.
H10 N5 avian influenza virus(AIV) of A/Duck/HuB/S1254/2014 was isolated from duck in a live poultry market in Hubei province during the routine epidemiologic surveillance in 2014. The virus complete genome was sequenced to know biological characteristics, the pathogenicity of the AIV was studied in BABL/c mice as a model, which was evaluated for mammalian infectivity. Sequence analysis of the hemagglutinin(HA) gene results showed that the virus was a characteristic of low pathogenic AIV, with the typical HA cleavage site of 334 PELMQGR↓GLF343. The highest nucleotide sequence of HA, NA, PA and PB2 genes were similar to A/migratoryduck/Jiangxi/30246/2013(H10 N5). However, the other internal gene segments of PB1, NP,M and NS genes originated from multiple subtypes. Pathogenicity experiments showed that the virus could be replicated in the turbinate and lung of mice without prior adaptation, with virus titers of 4.08 log10 EID50/mL and 5.0 log10 EID50/mL, respectively,The virus showed low pathogenicity to the mice, no obvious clinical symptoms and significant change of body weight during infection in mice. The results will contribute to understand the molecular evolutionary relationship of H10 N5 AIV and provide areference for evaluating the potential threat of H10 N5 AIV infection in mammals.
引文
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[2]Vijaykrishna D,Deng Y M,Su Y C,et al.The recent establishment of North American H10 lineage influenza viruses in Australian wild waterfowl and the evolution of Australian avian influenza viruses[J].J Virol,2013,87(18):10182-10189.
[3]Berg M,Englund L,Abusugra I A,et al.Close relationship between mink influenza(H10N4)and concomitantly circulating avian influenza viruses[J].Arch Virol,1990,113(1-2):61-71.
[4]Zhang Miao-miao,Zhang Xing-xing,Xu Kai-di,et al.Characterization of the pathogenesis of H10N3,H10N7,and H10N8subtype Avian influenza viruses circulating in ducks[J].Sci Rep,2016,6:34489.
[5]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.
[6]Wang Nan,Zou Wei,Yang Ying,et al.Complete genome sequence of an H10N5 avian influenza virus isolated from pigs in central China[J].J Virol,2012,86(24):13865-13866.
[7]Hoffmann E,Stech J,Guan Y,et al.Universal primer set for the full-length amplification of all influenza A viruses[J].Arch Virol,2001,146(12):2275-2289.
[8]Zhang Xu-xiao,Xu Guan-long,Wang Chen-xi,et al.Enhanced pathogenicity and neurotropism of mouse-adapted H10N7 influenza virus are mediated by novel PB2 and NA mutations[J].J Gen Virol,2017,98(6):1185-1195.
[9]Qi Xian,Li Xi-han,Rider P,et al.Molecular characterization of highly pathogenic H5N1 avian influenza A viruses isolated from raccoon dogs in China[J].PLo S One,2009,4(3):e4682.
[10]Wu Hai-bo,Lu Ru-feng,Wu Xiao-xin,et al.Novel reassortant H10N7 avian influenza viruses isolated from chickens in Eastern China[J].J Clin Virol,2015,65:58-61.
[11]Deng Guo-hua,Tan Dan,Shi Jian-zhong,et al.Complex reassortment of multiple subtypes of avian influenza viruses in domestic ducks at the Dongting Lake region of China[J].J Virol,2013,87(17):9452-9462
[12]Wood G W,Banks J,Strong I,et al.An avian influenza virus of H10 subtype that is highly pathogenic for chickens,but lacks multiple basic amino acids at the haemagglutinin cleavage site[J].Avian Pathol,1996,25(4):799-806.
[13]Jia Yane,Yang Jia-yun,Wang Zheng-xiang,et al.Genetic properties and pathogenicity of a novel reassortant H10N5 influenza virus from wild birds[J].Arch Virol,2017,162(5):1349-1353.
[14]Shi Jian-zhong,Deng Guo-hua,Kong Hui-hui,et al.H7N9 virulent mutants detected in chickens in China pose an increased threat to humans[J].Cell Res,2017,27(12):1409-1421.
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