A Rapid, Equipment-Free Method for Detecting Phytophthora infestans in the Field Using a Lateral Flow Strip-Based Recombinase Polymerase Amplification Assay
文献类型： 外文期刊
作者： Lu, Xinyu 1 ; Zheng, Ying 1 ; Zhang, Fan 1 ; Yu, Jia 1 ; Dai, Tingting 2 ; Wang, Rongbo 3 ; Tian, Yuee 4 ; Xu, Heng 1 ; Shen 1 ;
作者机构： 1.Nanjing Agr Univ, Coll Plant Protect, Nanjing 210095, Peoples R China
2.Nanjing Forestry Univ, Coll Forestry, Nanjing 210037, Peoples R China
3.Fujian Acad Agr Sci, Inst Plant Protect, Fuzhou 350013, Peoples R China
4.Henan Univ Sci & Technol, Dept Plant Protect, Luoyang 471000, Peoples R China
关键词： diagnostics; isothermal amplification; oomycetes; RPA; visualization
期刊名称：PLANT DISEASE （ 影响因子：4.438； 五年影响因子：4.7 ）
ISSN： 0191-2917
年卷期： 2020 年 104 卷 11 期
页码：
收录情况： SCI
摘要： Late blight, caused by the oomycete Phytophthora infestans, is a major constraint on the production of potatoes and tomatoes as well as a constant threat to global food security. An early diagnostic tool is important for the effective management of late blight in the field. Here, in combination with a simplified DNA extraction method, we developed a lateral flow strip-based recombinase polymerase amplification (LF-RPA) assay for the rapid, equipment-free detection of P. infestans. This assay targets the Ras-related protein (Ypt1) gene and can be performed over a wide range of temperatures (25 to 45 degrees C). All 12 P. infestans isolates yielded positive detection results using the LF-RPA assay, and no cross-reaction occurred with related oomycetes or fungal species. With this assay, the detection limit was 500 fg of genomic DNA in optimized conditions. Furthermore, by combining a simplified polyethylene glycol-NaOH method for extracting DNA from plant samples, the entire LF-RPA assay enabled the detection of P. infestans within 30 min with no specialized equipment. When applied to field samples, it successfully detected P. infestans in naturally diseased potato plants from eight different fields in China. Therefore, the LF-RPA assay is simple, rapid, and cost-effective and has potential for further development as a kit for diagnosing late blight in resource-limited settings or even on-site.
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