CAJ | 학술논문

　　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16S rRNA 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1 h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。
　　목적건립번가궤양병균적배개도등온핵산확증기술검측방법。방법응용배개도등온핵산확증기술(LAMP)대번가궤양병균적16S rRNA 특이성기인진행확증,채용4대인물식별목적편단적6개특이성구역,62℃항온1 h완성확증。결과설계적인물여대조균추문가단포균、가가단포균、정향가단포균번가치병변충도몰유확증반응,표현료교호적특이성。저사대조균재가과소채상역인기유사증상。결론 LAMP검측정서편첩,소수설비간단,기결과가이통과육안관찰래판단,비상규PCR령민차능경조득도반응결과,인차해기술구유경대적우세。
Objective To establish a method for detection of Clavibacter michiganensis subsp. Michiganensis by loop-mediated isothermal amplification protocol (LAMP). Methods Detection of 16S rRNA of Clavibacter mi-chiganensis subsp. Michiganensis by a loop-mediated isothermal amplification protocol (LAMP) was evaluated in laboratory assays. LAMP amplified target DNA rapidly (1 h), isothermally (62℃) and with high-specificity based on four primers designed to recognize six independent sequences of target DNA. Results The LAMP primers did not react with suspensions of Pseudomonas corrugata, Pseudomonas solanacearum, and Pseudomonas syringae pv. to-mato. The specificity of this LAMP assay was higher than PCR. Conclusion This method has convenient testing procedures, the required equipment is simple, and the results can be judged by the naked eye. It is sensitive than con-ventional PCR reaction and the results can be obtained earlier, so it has a greater advantage of the technology.