陕西省avian-like H1N1猪流感病毒的遗传进化分析

doi:10.3864/j.issn.0578-1752.2013.18.021

摘要/Abstract

摘要： 【目的】监测陕西省猪流感的流行与传播情况，为猪流感研究提供参考。【方法】从生猪养殖场和屠宰场表观健康猪中随机采取猪鼻/气管拭子样本，经SPF鸡胚分离，对HA阳性样本进行猪流感病毒RT-PCR鉴定。测定分离株全基因序列，通过MEGA4构建基因进化树，并进行遗传变异分析。【结果】从500份样本中分离鉴定出6株H1N1流感病毒，病毒分离率为1.2%。获得6个核苷酸相似性为98.3%—99.1%的分离株全基因序列。遗传进化分析表明6个分离株都位于avain-like H1N1猪流感病毒谱系，与中国江苏，辽宁，山东和香港等地H1N1猪流感病毒株同源性达97%—99%。6个分离株的8个基因片段均在蛋白受体结合位点、潜在糖基化位点等处发生核苷酸变异，这将会影响猪流感病毒的抗原特性、宿主适应性和致病力。【结论】首次从西北地区表观健康猪中分离到H1N1猪流感病毒，并获得6个分离株的全基因序列。遗传进化分析表明，6个分离株为avain-like H1N1猪流感病毒, 且多处氨基酸位点发生变异。

关键词: avain-like H1N1猪流感病毒 , 分离鉴定 , 遗传进化

Abstract: 【Objective】The prevalence and transmission of swine influenza in Shaanxi Province were studied.【Method】Nasal/tracheal swap samples were randomly collected from apparently healthy pigs in farms and slaughterhouses in Shaanxi province between October 2011 and May 2012. The samples were firstly separated by SPF chick embryos, and the HA-positive samples were identified as swine influenza virus using RT-PCR. Then, the full genomes of these isolates were sequenced and used to re-construct phylogenetic trees with software MEGA 4 and analyze genetic variations. 【Result】 Six H1N1 influenza viruses were isolated and identified among 500 samples, with isolation rate of 1.2%. The full length genomes were obtained from six isolates, with nucleotide similarity of 98.3–99.1% . Phylogenetic analysis showed that six H1N1 virus isolates were located into the lineage of avain-like H1N1 swine influenza viruses. The nucleotide similarities were 97–99% between six isolates and SIVs from Jiangsu, Liaoning, Shandong, and Hong Kong. The mutated nucleotides of eight gene fragments for six isolates were detected at receptor binding sites and potential glycosylation sites. These genetic variations will affect antigenicity, host adaptability and virulence of the viruses.【Conclusion】 The H1N1 swine influenza virus were firstly isolated from apparently healthy pigs in Northwestern China, and six full length genomes of these isolates were obtained. Phylogenetic analysis suggested six isolates were avain-like H1N1 swine influenza virus. Mutations were also detected in some amino acid sites of these isolates.

Key words: avain-like H1N1 SIV , isolation and identification , genetic evolution

任娟娟, 王晶钰, 刘红彦, 刘万华, 武宁, 邱渊皓, 唐攀, 江跃德. 陕西省avian-like H1N1猪流感病毒的遗传进化分析[J]. 中国农业科学, 2013, 46(18): 3901-3913.

REN Juan-Juan, WANG Jing-Yu, LIU Hong-Yan, LIU Wan-Hua, WU Ning, QIU Yuan-Hao, TANG Pan, JIANG Yue-De. Phylogenetic Analysis of Avain-like H1N1 Swine Influenza Virus in Shaanxi Province[J]. Scientia Agricultura Sinica, 2013, 46(18): 3901-3913.

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参考文献

[1]祁贤, 陆承平. 猪流感病毒进化方式及其流行特点. 微生物学报, 2009, 49(9): 1138-1145.

Qi X, Lu C P. Swine influenza virus: evolution mechanism and Epidemic characterization-A review. Acta Microbiologica Sinica, 2009, 49(9): 1138-1145. (in Chinese)

[2]Ito T, Couceiro J N, Kelm S, Baum L G, Krauss S. Castrucci M R, Donatelli I, Kida H, Paulson J C, Webster R G, Kawaoka Y. Molecular basis for the generation in pigs of Influenza A viruses with pandemic potential. Journal of Virology, 1998, 72: 7367-7373.

[3]Thacker E, Janke B. Swine influenza virus: zoonotic potential and vaccination strategies for the control of avian and swine influenzas. The Journey of Infectious Diseases, 2008, 197: S19--S24.

[4]Karasin A I, Carman S, Olsen C W. Identification of human H1N2 and human-swine reassortant H1N2 and H1N1 influenza A viruses among pigs in Ontario，Canada (2003 to 2005). Journal of Clinical Microbiology, 2006, 44: 1123-1126.

[5]Naffakh N, van der Werf S. April 2009: an outbreak of swine-origin influenza A(H1N1)virus with evidence for human-to-human transmission. Microbes and Infection , 2009, 11: 725-728．

[6]Dasco C C, Couch R B, Six H R, Young J F, Quarles J M, Kasel J A. Sporadic occurrence of zoonotic swine influenza virus infections. Journal of Clinical Microbiology, 1984, 20: 833-835.

[7]Wentworth D E, Thompson B L, Xu X, Reqnery H L, Cooley A J, McGregor M W, Cox N J, Hinshaw V S. An influenza virus A(H1N1) virus closely related to swine influenza responsible for a fetal case of human influenza. Journal of Virology, 1994, 68: 2051-2058.

[8]Vijaykrishna D, Poon L L, Zhu H C, Ma S K, Li O T, Cheung C L, Smith G J, Peiris J S, Guan, Y. Reassortment of pandemic H1N1/2009 influenza A virus in swine. Science, 2010, 328: 1529.

[9]Song M S , Lee J H, Pascua P N, Baek Y H, Kwon H I, Park K J, Choi H W, Shin Y K, Song, Y K, Kim C J, Choi Y K. Evidence of human-to-swine transmission of the pandemic (H1N1) 2009 influenza virus in South Korea. Journal of Clinical Microbiology, 2010, 48: 3204-3211.

[10]李海燕, 于康震, 辛晓光, 李雁冰, 蔡雪辉, 杨焕良, 毕英佐, 王自振．部分省市猪群猪流感的血清学调查及猪流感病毒的分离与鉴定. 动物医学进展, 2003, 24 (3): 67-72.

Li H Y, Yu K Z, Xin X G, Li Y B, Cai X H, Yang H L, Bi Y Z, Wang Z Z. Serological survey of swine influenza and isolation and identification of swine influenza virus in the herd of some provinces and cities. Progress in Veterinary Medicine, 2003, 24(3)：67-72. (in Chinese)

[11]吕翠, 马小明, 尹燕博, 单虎. H1亚型猪流感病毒的分离鉴定. 青岛农业大学学报, 2009, 26(1): 15-18.

Lü C, Ma X M, Yin Y B, Shan H. H1 Isolation and identification of H1 subtype swine influenza virus . Journal of Qingdao Agricultural University, 2009, 26 (1): 15-18. (in chinese)

[12]Hoffmann E, Stech J, Guan Y, Webster R G, Perez D R. Universal primer set for the full-length amplification of all influenza A viruses. Archives of Virology, 2001, 146 (12): 2275-2289.

[13]Tamura K, Dudley J, Nei M, Kumar S. MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Molecular Biology and Evolution , 2007, 24(8):1596-1599.

[14]Webster R G, Bean W J, Gorman O T, Chambers T M, Kawaoka Y. Evolution and ecology of influenza-A viruses. Microbiology Reviews , 1992, 56: 152-179.

[15]Matrosovich M, Tuzikov A, Bovin N, Gambaryan A, Klimov A, Castrucci M R, Donatelli I, Kawaoka Y. Early alterations of the receptor-binding properties of H1, H2, and H3 avian influenza virus hemagglutinins after their introduction into mammals. Journal of Virology, 2000, 74: 8501-8512.

[16]Igarashi M, Ito K, Kida H, Takada A. Genetically destined potentials for N-linked glycosylation of influenza virus hemagglutinin. Virology, 2008, 376: 323-329.

[17]Scholtissek C, Quack G, Klenk H D, Webster R G. How to overcome resistance of influenza Aviruses against adamantane derivatives. Antiviral Research, 1998, 37: 83-95.

[18]Suzuki H, Saito R, Masuda H, Oshitani H, Sato M, Sato I. Emergence of amantadine-resistant influenza Aviruses: epidemiological study. Journal of Infection and Chemotherapy, 2003, 9: 195-200.

[19]Peiris J S, de Jong, M D, Guan Y. Avian Influenza Virus (H5N1): a Threat to Human Health. Clinical Microbiology Reviews, 2007, 20: 243-267.

[20]Gabriel G, Abram M, Keiner B, Wagner R, Klenk H D, Stech J. Differential polymerase activity in avian and mammalian cells determines host range of influenza virus. Journal of Virology, 2007, 81: 9601-9604.

[21]Hatta M, Gao P, Halfmann P, Kawaoka Y. Molecular basis for high virulence of Hong Kong H5N1 influenza A viruses. Science, 2001, 293: 1840-1842.

[22]Labadie K, Dos Santos Afonso E, Rameix-Welti M A, van der Werf S, Naffakh N. Host-range determinants on the PB2 protein of influenza A viruses controlthe interaction between the viral polymerase andnucleoprotein in human cells. Virology, 2007, 362: 271-282.

[23]Subbarao E K, London W, Murphy B R. A single amino acid in the PB2 gene ofinfluenza A virus is a determinant of host range. Journal of Virology, 1993, 67: 1761-1764.

[24]Saito R, Sakai T, Sato I, Sano Y, Oshitani H, Sato M. Frequency of amantadine-resistant influenza A viruses during two seasons featuring cocirculation of H1N1 and H3N2. Journal of Clinical Microbiology, 2003, 41: 2164-2165.

[25]Shirashi K, Mitamura K, Sakai-Tagawa Y, Goto H, Sugaya N, Kawaoka Y. High frequency of resistant viruses harboring different mutation in amantadine-treated children with influenza. The Journey of Infectious Diseases, 2003, 188: 57-61.

[26]Shope R E. Swine Influenza : III. Filtration Experiments and Etiology. Journal of Experimental Medicine, 1931, 54(3): 373-385.

[27]Smith G J, Vijaykrishna D, Bahl J, Lycett S J, Worobey M, Pybus O G, Ma S K, Cheung C L, Raghwani J, Bhatt S, Peiris J S, Guan Y, Rambaut A. Origins and evolutionary genomics of the 2009 swine-origin H1N1 influenza A epidemic. Nature, 2009, 459: 1122-1125.

[28]Vijaykrishna D, Smith G J, Pybus O G, Zhu H, Bhatt S, Poon L L, Riley S, Bahl J, Ma S K, Cheung C L, Perera R A, Chen H. Long-term evolution and transmission dynamics of swine influenza A virus. Nature, 2011, 473: 519-522.

[29]Liu J H, Bi Y H, Qin K, Fu G H, Yang J, Peng J S, Ma G P, Liu Q F, Pu J, Tian F. Emergence of European avian influenza virus-like H1N1 swine influenza A viruses in China. Journal of Clinical Microbiology, 2009, 47: 2643-2646.

[30]Yu H, Zhang P C, Zhou Y J, Li G X, Pan J, Yan L P, Shi X X, Liu H L, Tong G Z. Isolation and genetic characterization of avian-like H1N1 and novel ressortant H1N2 influenza viruses from pigs in China. Biochemical and Biophysical Research Communications, 2009, 386: 278-283.

[31]Qi X, Pan Y, Qin Y, Zu R, Tang F, Zhou M, Wang H, Song Y. Molecular characterization of avian-like H1N1 swine influenza a viruses isolated in Eastern China, 2011. Virologica Sinica, 2012, 27(5): 292-298.

[32]Rimmelzwaan G F, de Jong J C, Bestebroer T M, van Loon A M, Claas E C, Fouchier R A , Osterehaus A D. Antigenic and genetic characterization of swine influenza A (H1N1) viruses isolated from pneumonia patients in the Netherlands. Virology, 2001, 282: 301-306.

[33]de Jong J C, Claas E C, Osterhaus A D, Webster R G, Lim W L. A pandemic warning? Nature, 1997, 389: 554.

[34]Myers K P, Olsen C W, Gray G C. Cases of swine influenza in humans:a review of the literature. Clinical Infectious Diseases, 2007, 44(8): 1084-1088.

[35]Wang J Y, Ren J J, Qiu Y H, Liu H J. Complete genome sequences of six avian-like H1N1 swine influenza viruses from northwestern China. Genome Announcement, 2013, 1(1) :e00098-12.