Jiang Junye, Feindel Will, Harding Michael, Feindel David, Bajema Stacey, Feng Jie
Potato Growers of Alberta, Edmonton, AB, T5Y 6H3, Canada.
Crop Diversification Centre South, Alberta Agriculture, Forestry and Rural Economic Development (AAFRED), Brooks, AB, T1R 1E6, Canada.
Plant Dis. 2023 Jan;107(1):136-141. doi: 10.1094/PDIS-05-22-1000-SR. Epub 2023 Jan 9.
is the causal agent of powdery scab of potato (), which can significantly reduce potato quality. In this study, we developed and evaluated a loop-mediated isothermal amplification (LAMP) method for the detection of . A set of LAMP primers named PS-LAMP was designed and tested for specificity and sensitivity. In the specificity test, in silico analysis using the NCBI Primer-BLAST tool indicated that PS-LAMP was specific to . The in vitro tests confirmed specificity, showing that PS-LAMP could produce positive signals from DNA isolated from each of three potato tubers with powdery scab symptoms but did not produce positive signals from DNA isolated from 38 nontarget plant pathogens. The sensitivity of PS-LAMP was tested on both gBlocks and DNA isolated from potato samples with powdery scab symptoms. On gBlocks, the lowest number of copies for a positive LAMP reaction was six, which was similar to results obtained via qPCR, but it was 10 times more sensitive than conventional PCR. On a DNA sample from -infected potato, the lowest amount of template DNA for a positive LAMP reaction was 2 pg, which was incomparable with the sensitivity of qPCR. Considering the convenience of the LAMP technique, as well as the high specificity and sensitivity, this assay can be very useful for plant pathology practitioners and diagnostic labs interested in rapid, accurate, and routine detection of and confirmation of powdery scab disease.
是马铃薯粉痂病的致病因子（），可显著降低马铃薯品质。在本研究中，我们开发并评估了一种用于检测的环介导等温扩增（LAMP）方法。设计了一组名为PS-LAMP的LAMP引物，并对其特异性和灵敏度进行了测试。在特异性测试中，使用NCBI Primer-BLAST工具进行的电子分析表明PS-LAMP对具有特异性。体外测试证实了其特异性，表明PS-LAMP可从三个有粉痂病症状的马铃薯块茎中分离的DNA产生阳性信号，但从38种非靶标植物病原体中分离的DNA未产生阳性信号。在gBlocks和从有粉痂病症状的马铃薯样品中分离的DNA上测试了PS-LAMP的灵敏度。在gBlocks上，LAMP阳性反应的最低拷贝数为6个，这与通过qPCR获得的结果相似，但比传统PCR灵敏10倍。在来自感染马铃薯的DNA样品上，LAMP阳性反应的最低模板DNA量为2 pg，这与qPCR的灵敏度无可比性。考虑到LAMP技术的便利性以及高特异性和灵敏度，该检测方法对于对快速、准确和常规检测以及粉痂病确诊感兴趣的植物病理学从业者和诊断实验室可能非常有用。
