果生刺盘孢菌LFD-RPA可视化快速检测方法的建立

The Visual Rapid Detection of Colletotrichum fructicola Based on LFD-RPA Assay

建立针对果生刺盘孢菌(Colletotrichum fructicola)的快速、准确、简便的检测方法。根据果生刺盘孢菌的基因序列设计了特异性检测引物和探针,建立了重组酶聚合酶扩增与侧流层析试纸条(LFD-RPA)相结合的可视化快速检测果生刺盘孢菌的方法。该检测方法与其他病原菌无交叉反应,特异性强,且检测灵敏度达1 pg/μL。该检测方法无需PCR仪等复杂设备,在39℃恒温反应12 min即可完成检测过程,为果生刺盘孢菌的检验检测和早期预警提供了技术支撑。

In order to establish a rapid,accurate and simple detection method of Colletotrichum fructicola.A visual rapid detection assay of C.fructicola was established by designing specific primers and probe according to the gene sequence of C.fructicola based on the combination recombinase polymerase amplification with lateral flow dipsticks(LFD-RPA).The detection method had strong specificity and high sensitivity(1 pg/μL),besides,there was no cross-reaction with other pathogens.This method could be applied without any expensive and complex equipments such as PCR apparatus,meanwhile,the temperature of reaction condition was 39℃and detection time was less than 12 min.This study provided a quick and simple technology for the detection and early warning of Colletotrichum fructicola.