›› 2010, Vol. 37 ›› Issue (6): 78-83.

• 生物技术 • 上一篇    下一篇

猪流感病毒的分离及NA基因的遗传进化分析

侯韶毅1,冯建远1,高永锐2,卢桂娟2,陆有飞2,肖爱欢1,张民秀1,卢冰霞1,黄伟坚1,赵武3   

  1. (1.广西大学动物科学技术学院, 南宁 530005; 2.广西丽原生物股份有限公司, 南宁 530001;3.广西兽医研究所, 南宁 530001)
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2010-06-20 发布日期:2010-06-20
  • 通讯作者: 黄伟坚

Isolation of Swine Influenza A Viruses and Genetic Characterization for NA Genes of the Isolated Strains

HOU Shao-yi1, FENG Jian-yuan1, GAO Yong-rui2, LU Gui-juan2, LU You-fei2, XIAO Ai-huan1,
ZHANG Min-xiu1, LU Bing-xia1, HUANG Wei-jian1, ZHAO Wu3
  

  1. (1.College of Animal Science and Technology, Guangxi University, Nanning 530005, China; 2.Guangxi Liyuan Biologic CoLtd,Nanning 530001,China; 3.Guangxi Veterinary Research Institute, Nanning 530001, China)
  • Received:1900-01-01 Revised:1900-01-01 Online:2010-06-20 Published:2010-06-20
  • Contact: HUANG Wei-jian

摘要: 通过鸡胚接种分离到3株猪流感病毒,经HI、RT-PCR鉴定,其中GXHZ株为H1N2亚型毒株,GXLZ、GXXY株为H3N2亚型毒株,并对分离株进行EID50测定及GXHZ株的猪体攻毒试验。所扩增分离到3株SIV病毒NA基因与A/Sw/Hainan/1/2005(H1N2)的核苷酸和氨基酸序列同源性最高。3株SIV分离株的NA基因在氨基端胞浆尾区均由K6→R6,在非极性跨膜区均由V20→M20。系统发育分析表明本试验所分离的3个SIV毒株与H1N2亚型毒株亲缘性最相近,且来源于猪源H1N2亚型流感病毒。从时间上也发现,3个分离株的NA基因与1996年以后获得的H3N2毒株亲缘关系很近,都隶属于一个亚分支。

关键词: 猪流感病毒; 病毒分离; NA基因; 遗传进化分析

Abstract: Three swine influenza virus strains were isolated from 9-11-day SPF chicken eggs in our study of these, GXHZ isolation was subtyped as H1N2 by HI assay and RT-PCR, and GXLZ,GXXY as H3N2. We detected the EID50 of these isolated strains and performed inoculation to pig with GXHZ isolation. NA genes amplificated from 3 isolated strains showed that their nucleotide and amino acid sequences had highest homology to A/Sw/Hainan/1/2005 (H1N2) strain. Two substitution at 6 (K6→R6) and 20 (V20→M20) separately located on the amino acid cytoplasmic tail and nonpolar transmembrane domain of the NA molecules of isolated strains. Phylogenical analysis of the NA genes indicated that the NA genes of isolated strains were most closely related to H1N2 which derived from swine origin influenza virus. Moreover, three isolated strains were clustered with H3N2 strains isolated after 1996.

Key words: swine influenza virus; viral isolation; NA gene; analysis of genetic evolution

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