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中国农学通报

中国农学通报 ›› 2012, Vol. 28 ›› Issue (20): 83-87.doi: 10.11924/j.issn.1000-6850.2011-3379

所属专题： 畜牧兽医

• 畜牧 动物医学 蚕 蜂 • 上一篇    下一篇

鸡传染性支气管炎病毒RT-LAMP可视化检测方法的建立

罗思思 谢芝勋 庞耀珊 谢志勤 邓显文 刘加波 谢丽基 彭宜 范晴   

  • 收稿日期:2011-11-15 修回日期:2011-11-29 出版日期:2012-07-15 发布日期:2012-07-15

Development of a Reverse Transcription Loop-mediated Isothermal Amplification Assay

  • Received:2011-11-15 Revised:2011-11-29 Online:2012-07-15 Published:2012-07-15

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摘要：

为建立一种能快速检测鸡传染性支气管炎病毒(IBV)的方法，根据GenBank中IBV的基因保守序列，设计一套环介导等温扩增(LAMP)特异性引物，建立了IBV RT-LAMP可视化检测方法；该法对IBV RNA最小检测限为10 fg，而常规RT-PCR为1 pg，灵敏度高于常规PCR法100倍；对其他常见鸡病原体检测结果均为阴性；可通过肉眼观察颜色直接判定结果；本研究建立的IBV RT-LAMP方法简便、快捷、特异、灵敏，且只需1个可控温的水浴锅在1 h内即可完成全部反应，更适合基层业务部门及养殖场的检测，为IBV感染的快速检测提供新方法。

关键词: 运行机制, 运行机制

Abstract:

A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for detection of infectious bronchitis virus(IBV); According to the sequences of IBV in GenBank, six primers were designed, and the reaction conditions were optimized. The results showed that the detection limit of this RT-LAMP method was 10 fg, whereas that of RT-PCR was 1 pg, suggesting the LAMP method was much more sensitive than the RT-PCR methods for IBV detection, the LAMP method for IBV detection was more convenient, rapid, specific and sensitive than the RT-PCR, the assay was performed in water bath within 1 h. Therefore, it was very suitable for detection in the field condition that would provide a novel rapid detection approach for IBV infection.

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