华北农学报 ›› 2015, Vol. 30 ›› Issue (S1): 342-347. doi: 10.7668/hbnxb.2015.S1.062

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猪流行性腹泻病毒TX株的分离鉴定及毒力分析

于晓明, 陈瑾, 侯立婷, 吴楠, 侯继波, 郑其升   

  1. 国家兽用生物制品工程技术研究中心, 江苏 南京 210014
  • 收稿日期:2015-10-11 出版日期:2015-12-28
  • 通讯作者: 郑其升(1979-),男,山东潍坊人,副研究员,博士,主要从事猪病新型疫苗研究.
  • 作者简介:于晓明(1985-),男,山东威海人,助理研究员,硕士,主要从事猪流行性腹泻病毒疫苗研究.
  • 基金资助:
    江苏省农业自主创新专项(CX(14)2089)

Isolation,Identification and Virulence Analysis of Porcine Epidemic Diarrhea Virus TX Strain

YU Xiao-ming, CHEN Jin, HOU Li-ting, WU Nan, HOU Ji-bo, ZHENG Qi-sheng   

  1. National Research Center of Veterinary Biological Engineering and Technology, Nanjing 210014, China
  • Received:2015-10-11 Published:2015-12-28

摘要: 2011年江苏泰兴某些猪场相继发生了以腹泻为主要症状的疾病,造成大量哺乳仔猪甚至育肥猪发生死亡.对采集腹泻病料进行病毒的分离鉴定及毒力分析.首先用特异性引物对采集的病料样品进行常见猪病的PCR或RT-PCR检测,并处理病料组织进行电镜观察;腹泻病毒经1日龄未吃初乳的仔猪进行猪体复归试验,再次收取病料并进行病毒纯净性检测;最后测定了腹泻组织毒对5,15,25日龄仔猪的致病性,及对25日龄仔猪的半数致死量.结果表明:PCR与RT-PCR鉴定结果腹泻猪为猪流行性腹泻病毒感染所致,在组织中没有其他常见猪传染病病原的存在;电镜观察呈典型的PEDV冠状形态;以特异性引物扩增 S1 基因,分析测序结果并进行基因进化树分析表明该毒株与当前流行性腹泻疫苗株亲缘关系较远,而与我国当前流行的PEDV变异株亲缘关系近;该毒株对5,15,25日龄仔猪100%致死,感染后表现腹泻、呕吐等典型的PEDV感染症状.

关键词: 猪流行性腹泻病毒, 分离鉴定, 进化分析, 致病力

Abstract: Porcine epidemic diahorrea virus (PEDV) TX strain was isolated from the feces of piglets suspected to be suffering from a severe diarrhea in Jiangsu Province.To investigate the occurrence and origin of diarrhea in piglet,porcine epidemic disease virus (PEDV),transmissible gastroenteritis coronavirus (TGEV) and porcine rotavirus (RV) et al were detected by RT-PCR.Typical coronavirus particles were observed by transmission electron microscopy(TEM) in feces samples.The phylogenetic tree was established by comparing the S1 gene amplifying from TX strain with other PEDV strains.In order to identified the virus pathogenicity,age of 5,15,25 days piglet were challenged with TX strain.The samples were only positive to PEDV detected by RT-PCR.The TX strain shared a highly homology of 94.8%-99.3% with epidemic strains.All the piglets died after were challenged with virus,indicating a high pathogenicity.

Key words: Porcine epidemic diarrhea virus, Isolation and identification, Analysis of phylogenetic, Virulence

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引用本文

于晓明, 陈瑾, 侯立婷, 吴楠, 侯继波, 郑其升. 猪流行性腹泻病毒TX株的分离鉴定及毒力分析[J]. 华北农学报, 2015, 30(S1): 342-347. doi: 10.7668/hbnxb.2015.S1.062.

YU Xiao-ming, CHEN Jin, HOU Li-ting, WU Nan, HOU Ji-bo, ZHENG Qi-sheng. Isolation,Identification and Virulence Analysis of Porcine Epidemic Diarrhea Virus TX Strain[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2015, 30(S1): 342-347. doi: 10.7668/hbnxb.2015.S1.062.

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