烟草靶斑病菌LFD-RAA快速检测方法的建立OA北大核心CSTPCD
A rapid LFD-RAA detection method for tobacco target spot pathogen
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.
To rapidly and effectively detect the tobacco target spot pathogen,the conserved region of the beta-tubulin gene sequence of the pathogen was targeted and the recombinase aided amplification(RAA)primers were designed.Lateral flow chromatographic strips were used to establish and further optimize a rapid detection system.The results showed that the developed RAA amplification could be completed within 30 minutes at a constant temperature of 39 ℃ and had high specificity with a detection limit of 100 fg/μL.Its sensitivity was comparable to that of conventional PCR detections.The optimal amplification time and temperature of the optimized detection system were 20-35 min and 31-39 ℃.This LFD-RAA detection system could be applied for rapid detection of the pathogen of tobacco target spot on tobacco plants.
肖艳松;李思军;吴文信;刘天波;周向平;周陆苏;钟杰
湖南省烟草公司郴州市公司,湖南省郴州市北湖区燕泉街道燕泉北路61号 423000湖南省烟草科学研究所,长沙市芙蓉南路368号 410004湖南省烟草公司永州市公司,湖南省永州市冷水滩区珍珠北路69号 425000湖南农业大学植物保护学院,长沙市芙蓉区农大路1号 410128
植物保护学
烟草靶斑病重组酶介导扩增技术检测
Tobacco target spotRecombinase aided amplificationDetection
《烟草科技》 2024 (005)
41-47 / 7
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