摘要：为快速有效检测烟草靶斑病菌,以烟草靶斑病菌beta-tubulin基因序列保守区域为靶标位点,设计重组酶介导扩增技术(Recombinase aided amplification,RAA)引物,结合侧流层析试纸条(Lateral flow dipstick,LFD),建立烟草靶斑病菌快速检测体系并进行优化。结果表明,LFD-RAA检测体系在39℃恒温条件下反应30 min即可完成RAA扩增,并具有较高的特异性,其检测极限值为100 fg/μL,与PCR检测灵敏度相当。对检测体系优化确定检测扩增的最优时间为20~35 min、扩增温度为31~39℃。LFD-RAA检测体系可应用于烟草上烟草靶斑病菌的快速检测。

摘要：This study aimed to achieve rapid detection of Pseudomonas syringae ***,the pathogen of tobacco wildfire *** specific primers and probes for recombinase-aided amplification(RAA)were designed with HrpZ as the target *** was then combined with the lateral flow dipstick(LFD)to establish a LFD-RAA-based rapid detection system for the ***,the detection performance of the established method was *** results showed that the LFDRAA method had high *** amplification could be completed after 25 min of reaction at 39℃.The sensitivity of the established method reached 0.0001 ng/μL,which was superior to that of PCR ***,the LFD-RAA method could quickly detect *** *** from tobacco leaves,demonstrating field *** sum up,the LFD-RAA method established in this study can be applied in the rapid detection and early diagnosis of tobacco wildfire disease.