环介导等温扩增技术检测菊花中番茄不孕病毒

【目的】环介导等温扩增(loop-mediated isothermal amplification,LAMP)是一种特异、灵敏、快速的新型基因检测技术。本研究拟用该技术建立菊花中番茄不孕病毒(TAV)的检测方法。【方法】设计能识别TAV CP序列6个特定区域的4种LAMP特异引物,对MgCl2、dNTPs、甜菜碱等LAMP反应条件进行优化,利用新建的LAMP法检测TAV,比较LAMP和RT-PCR2种不同方法的检测灵敏度。【结果】建立了检测番茄不孕病毒的LAMP技术;LAMP比RT-PCR具有灵敏度更高、快速、简便等优点。【结论】建立的LAMP技术能快速、准确、简便地检测出菊花中的番茄不孕病毒。

Detection of Tomato Aspermy Virus Infecting Chrysanthemums by LAMP

Objective]In this study,the LAMP (loop-mediated isothermal amplification) was used to detect TAV (Tomato aspermy virus). Method]Using TAV CP cDNA clones as templates,the reaction conditions (such as Mg~(2+),dNTPs,betaine and primers concentrations) optimized and the sensitivities of LAMP and RT-PCR were compared in detecting TAV. Result]The LAMP method was developed in this experiment and it was more sensitive than RT-PCR method for detecting TAV. Conclusion]The LAMP as a simple and quick method can accurately detect TAV in chrysanthemum samples.