中国农业科学 ›› 2006, Vol. 39 ›› Issue (01): 215- .

• 研究简报 • 上一篇    

SYBR GreenI模式荧光RT-PCR鉴别新城疫病毒毒力

弋英,陈继明,王志亮,孙承英,鲍恩东   

  1. 南京农业大学动物医学院
  • 收稿日期:2004-07-26 修回日期:2005-04-30 出版日期:2006-01-10 发布日期:2006-01-10
  • 通讯作者: 鲍恩东

Pathotyping of Newcastle Disease Virus by SYBR Green I Real-Time RT-PCR

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  1. 南京农业大学动物医学院
  • Received:2004-07-26 Revised:2005-04-30 Online:2006-01-10 Published:2006-01-10

摘要: 【方法】根据NDV的F蛋白裂解位点附近的序列,设计分别对应于NDV强毒和弱毒的两对引物,然后用SYBR Green I模式的荧光RT-PCR方法比较两对引物对同一NDV的RNA扩增效率以及扩增产物的Tm值,来区分NDV的毒力。【结果】对8株NDV进行检测并测定其鸡胚平均致死时间(MDT),荧光RT-PCR方法的检测结果和MDT测定的结果完全一致,说明此荧光RT-PCR法可用于NDV毒力检测。

关键词: 鸡, 新城疫病毒, SYBR Green I模式荧光RT-PCR, 毒力

Abstract: 【Method】The virulence of Newcastale disease virus (NDV) strains was determined by the lysis site sequences of the viral F protein. Accordingly, two pairs of primers were designed to match such sequences of virulent NDV strains and avirulent strains, respectively. Viral RNA was amplified using the two primers respectively by one-step SYBR Green I RT-PCR, and the virus was pathotypied with the amplification effects of the two pairs of primers and the Tm values of the amplification products. 【Conclusion】Pathotyping results of 8 unknown strains of this assay were as the same as those of Mean Death Time (MDT) and sequencing, which suggested that this assay could be used to pathotype NDV strains.

Key words: Avian, Newcastle disease virus, SYBR Green I Real-time RT-PCR, Pathotype