Aiying Wang, Ju Luo, Cilin Wang, Yuxuan Hou, Baojun Yang, Jian Tang, Shuhua Liu
State Key Laboratory of Rice Biology and Breeding, China National Rice Research Institute, Hangzhou, 310006 China.
Australas Plant Pathol. 2023 May 18:1-9. doi: 10.1007/s13313-023-00918-8.
is a major pathogen that causes the new bacterial blight in rice, and its symptoms very similar to rice bacterial blight. Therefore, there is a dire need for an accurate and rapid method for detecting . In this study, an early and rapid visual detection method for was established. Using GyrB gene as the target sequence, an innovative recombinase-aided amplification detection system integrated with a lateral flow dipstick (RAA-LFD) was constructed. The optimized RAA-LFD detection method can be initiated at body temperature and does not rely on precise instruments. It does not require DNA extraction and can be used directly with plant tissue fluids. The results can be visualized after 10 minutes of amplification. The specificity and sensitivity tests showed that the RAA-LFD method could detect whereas other common plant pathogens were not detected, and its detection sensitivity for DNA reached 100 copies/µL. The detection of diseased tissues indicated that this method could accurately detect in artificially inoculated rice tissues in the early stages of infection before symptoms. The RAA-LFD detection system established in this study is simple and fast, with visual results, excellent specificity, and high sensitivity. It is semi-quantitative and should be used for the early detection and rapid field diagnosis of new leaf blight, which provides technical support for the early warning and real-time detection of field samples.
是引起水稻新细菌性条斑病的主要病原菌,其症状与水稻白叶枯病非常相似。因此,迫切需要一种准确、快速的检测方法。本研究建立了一种早期快速的可视化检测方法。以GyrB基因作为靶序列,构建了一种与侧向流动试纸条相结合的创新重组酶辅助扩增检测系统(RAA-LFD)。优化后的RAA-LFD检测方法可在体温下启动,不依赖精密仪器。它不需要DNA提取,可直接用于植物组织液。扩增10分钟后即可观察到结果。特异性和敏感性测试表明,RAA-LFD方法能检测到,而未检测到其他常见植物病原菌,其对DNA的检测灵敏度达到100拷贝/µL。对病组织的检测表明,该方法能在症状出现前的感染早期准确检测人工接种水稻组织中的。本研究建立的RAA-LFD检测系统简单快速,结果可视化,特异性好,灵敏度高。它具有半定量功能,应用于新叶枯病的早期检测和田间快速诊断,为田间样本的预警和实时检测提供技术支持。