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烟草野火病菌LFD-RAA快速检测方法的建立

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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。
A LFD-RAA-Based Rapid Detection Method for Pseudomonas syringae pv.tabaci
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.

Tobacco wildfire diseaseRecombinase-aided amplificationDetection

肖艳松、李宏光、李思军、吴文信、周陆苏、钟杰、苏家恩、杨志娟

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湖南省烟草公司郴州市公司,湖南 郴州 423000

湖南省烟草公司郴州市公司桂阳县公司,湖南 郴州 424400

湖南农业大学植物保护学院,湖南 长沙 410128

云南省烟草公司大理市公司,云南 大理 671000

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烟草野火病 重组酶介导扩增技术 检测

2024

农业科学与技术(英文版)
湖南省农业科学院

农业科学与技术(英文版)

影响因子:0.222
ISSN:1009-4229
年,卷(期):2024.25(2)