Development of a real-time fluorescence loop-mediated isothermal amplification assay for rapid and quantitative detection of Fusarium mangiferae associated with mango malformation
文献类型： 外文期刊
作者： Pu, Jinji 1 ; Xie, Yixian 1 ; Zhang, He 1 ; Zhang, Xin 1 ; Qi, Yanxiang 1 ; Peng, Jun 1 ;
作者机构： 1.Chinese Acad Trop Agr Sci, Environm & Plant Protect Inst, Key Lab Integrated Pest Management Trop Crops, Minist Agr, Haikou 571101, Peoples R China
2.China Agr Univ, State Key Lab Agrobiotechnol, Beijing 100193, Peoples R China
3.China Agr Univ, Minist Agr, Key Lab Plant Pathol, B
关键词： Fusarium mangiferae;Mango malformation disease;Real-time fluorescence loop-mediated isothermal amplification assay (RealAmp);Quantitative detection;Closed-tube visual detection
期刊名称：PHYSIOLOGICAL AND MOLECULAR PLANT PATHOLOGY （ 影响因子：2.747； 五年影响因子：2.388 ）
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收录情况： SCI
摘要： Fusarium mangiferae is a major causal agent of mango malformation disease (MMD) worldwide. Rapid and accurate detection of the causal pathogen is the cornerstone of integrated disease management. In this paper, a real-time fluorescence loop-mediated isothermal amplification assay (RealAmp) was developed for quantitative detection of E mangiferae in China. The LAMP primer set was designed based on a RAPD marker sequence and positive products were amplified only from F. mangiferae isolates, but not from any other species tested, showing a high specificity of the primer sets. The detection limit was approximately 2.26 x 10(-4) ng/mu l plasmid DNA when mixed with extracted mango DNA. Quantification of the pathogen DNA of MMD in naturally collected samples was no significant difference compared to classic real-time PCR Additionally, RealAmp assay was visual with an improved closed-tube visual detection system making the assay more convenient in diagnostics.
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