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目的建立番茄溃疡病菌的环介导等温核酸扩增技术检测方法。方法应用环介导等温核酸扩增技术(LAMP)对番茄溃疡病菌的16SrRNA特异性基因进行扩增,采用4对引物识别目的片段的6个特异性区域,62℃恒温1h完成扩增。结果设计的引物与对照菌皱纹假单胞菌、茄假单胞菌、丁香假单胞菌番茄致病变种都没有扩增反应,表现了较好的特异性。这些对照菌在茄科蔬菜上易引起类似症状。结论 LAMP检测程序便捷,所需设备简单,其结果可以通过肉眼观察来判断,比常规PCR灵敏且能更早得到反应结果,因此该技术具有更大的优势。
Objective To establish a method for detecting ring-mediated isothermal nucleic acid amplification of tomato canker. Methods 16S rRNA specific genes of tomato canker were amplified by ring-mediated isothermal amplification of nucleic acid (LAMP). Four primers were used to amplify 16S rRNA specific primers and identified by PCR at 62 ℃ for 1 h. Results There was no amplification reaction between the primers designed and the control strains Pseudomonas putida, Pseudomonas solanacearum and Pseudomonas syringae pv. Tomato, which showed good specificity. These control bacteria easily cause similar symptoms in Solanaceae vegetables. Conclusion The LAMP detection procedure is simple, the equipment required is simple, the results can be judged by visual observation, which is more sensitive than conventional PCR and can get the reaction results earlier. Therefore, this technique has more advantages.