摘要
为快速有效检测烟草野火病菌(Ps-eudomonas syringae pv.tabaci),以烟草野火病菌HrpZ基因序列为靶标,设计重组酶介导扩增技术(recombinase aided amplification,RAA)特异性引物,结合侧流层析试纸条,获得了特异性的引物和探针,建立了烟草野火病菌LFD-RAA快速检测体系,并对其检测能力进行验证.结果表明:该检测体系具有较高的特异性,在 39℃恒温条件下反应 25 min即可完成扩增,灵敏度可达到 0.0001 ng/μL,优于PCR检测灵敏度,且该检测体系可从烟草叶片上快速检测到烟草野火病菌,证明其实用性强.综上所述,LFD-RAA检测体系可应用于烟草野火病的快速检测和早期诊断.
Abstract
This study aimed to achieve rapid detection of Pseudomonas syringae pv.tabaci,the pathogen of tobacco wildfire disease.The specific primers and probes for recombinase-aided amplification(RAA)were designed with HrpZ as the target gene.RAA was then combined with the lateral flow dipstick(LFD)to establish a LFD-RAA-based rapid detection system for the pathogen.Furthermore,the detection performance of the established method was tested.The results showed that the LFD-RAA method had high specificity.The 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 detection.Moreover,the LFD-RAA method could quickly detect P.syringae pv.tabaci from tobacco leaves,demonstrating field applicability.To sum up,the LFD-RAA method established in this study can be applied in the rapid detection and early diagnosis of tobacco wildfire disease.
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