欢迎访问《中国农学通报》，
中国农学通报

中国农学通报 ›› 2012, Vol. 28 ›› Issue (31): 206-210.

• 林学 园艺 园林 • 上一篇    下一篇

水仙花叶病毒RT-PCR检测方法的建立及应用

沈建国 高芳銮 蔡伟 于文涛 虞赟   

  • 收稿日期:2012-08-13 修回日期:2012-09-14 出版日期:2012-11-05 发布日期:2012-11-05
  • 基金资助:

    福建水仙病毒病的种类鉴定及新型快速分子诊断试剂盒研制

Establishment and Application of RT- PCR for detection of Narcissus mosaic virus

  • Received:2012-08-13 Revised:2012-09-14 Online:2012-11-05 Published:2012-11-05

PDF (PC)

825

摘要：

旨在建立水仙花叶病毒(Narcissus mosaic virus，NMV)的分子快速检测方法。根据已公布的NMV基因序列，设计一对特异性引物，以提取的水仙样品总RNA为模板进行RT-PCR，并对该方法的特异性、灵敏度、重复性及实际检测效果进行测定。从感染NMV的水仙样品中成功扩增出预期大小（约483 kb）的特异性目的片段，而健康水仙上未扩增到相应的片段。特异性、灵敏度及重复性测定结果表明，本研究建立的RT-PCR方法特异性强、灵敏高、重复性好。应用该方法成功检测了一批口岸送检的水仙样品。本研究建立的RT-PCR方法可为水仙上NMV的检测提供可靠的技术支持。

关键词: 高效液相色谱, 高效液相色谱

Abstract:

The aim was to establish a rapid molecular assay for detection of Narcissus mosaic virus (NMV). A pair of specific primers was designed according to the published nucleotide sequence of NMV. The cDNA was synthesized from the total RNA prepared from narcissus sample and then PCR was carried out. The specificity, sensitivity, repeatability and practical effect of detection were validated. The method could specifically amplify an expected 483 bp fragment from NMV-infected narcissus sample, but not from healthy narcissus sample. The results showed that the method had strong specificity, high sensitivity and good repeatability. The method was successfully used to detect NMV on narcissus samples from port. It was concluded that RT-PCR provided reliable technical support for detection of NMV in Narcissus.