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中国农学通报

中国农学通报 ›› 2012, Vol. 28 ›› Issue (29): 79-82.

• 畜牧 动物医学 蚕 蜂 • 上一篇    下一篇

水貂绿脓杆菌环介导等温扩增快速检测方法的建立

姜莉莉 樊兆斌 高文玉   

  • 收稿日期:2012-07-15 修回日期:2012-08-04 出版日期:2012-10-15 发布日期:2012-10-15
  • 基金资助:

    辽宁省教育厅基金项目

Rapid Detection for Mink Pseudomonas aeruginosa by Loop-mediated Isothermal Amplification

  • Received:2012-07-15 Revised:2012-08-04 Online:2012-10-15 Published:2012-10-15

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摘要：

为了利用环介导等温扩增(LAMP)技术建立水貂绿脓杆菌核酸快速检测方法。根据绿脓杆菌外毒素A基因240 bp片段保守序列设计一套LAMP特异性引物，对水貂绿脓杆菌基因组DNA进行LAMP扩增，通过浑浊度、SYBR Green荧光染料显色及电泳观察结果，并进行特异性和敏感性检验。结果表明：完成反应仅需要60 min(65℃)，特异性强，最低检测限为1 ng/μL。该方法简单快捷、特异性强、灵敏度高，适合于基层使用，具有广泛的临床应用前景。

关键词: 抗体, 抗体

Abstract:

In order to establish a rapid detection technique for mink Pseudomonas aeruginosa by Loop-mediated isothermal amplification. A set of specific primers were designed based on conserved region of PEA gene (Pseudomonas aeruginosa 240 bp), and amplified DNA for mink Pseudomonas aeruginosa by LAMP. The results were observed by turbidity, SYBR Green staining and agarose gel electrophoresis, meanwhile specificity and sensibility experiments were done too. The results showed that reaction was finished in 60 min, showed a high specificity and the detection limit was 1 ng/μL. The techniques were simple, rapid, high specificity and sensibility, so it could applied to base course and had comprehensive clinical prospect.