Genomics-informed design of loop-mediated isothermal amplification for detection of phytopathogenic Xanthomonas arboricola pv. pruni at the intraspecific level
Genomics-informed design of loop-mediated isothermal amplification for detection of phytopathogenic Xanthomonas arboricola pv. pruni at the intraspecific level
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DOI：
10.1111/j.1365-3059.2012.02654.x
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发表时间：
2013-04-01
期刊：
影响因子：
2.7
通讯作者：
Duffy, B.
中科院分区：
文献类型：
--
作者：
Buehlmann, A.;Pothier, J. F.;Duffy, B.
本研究的目的是建立一种快速、灵敏的检测方法，用于检测李属果树核果细菌性斑点病的病原菌--树生黄单胞菌李致病变种（Xanthomonasarboricolapv.pruni）。使用其他黄单胞菌属物种、亚种和致病变种的比较基因组学管道从树栖黄单胞菌李致病变种基因组鉴定独特靶标，并用于鉴定特异性诊断标记。环介导等温扩增（LAMP），然后应用于这些标志物，以提供快速，灵敏和特异性的检测。开发的方法显示出无与伦比的特异性与79个测试菌株，并在以前建立的技术相比，区分与遗传学密切的亚种，如X.arboricola pv. corylina。该测试的灵敏度与先前报道的TaqMan测定在103 CFUmL 1下的灵敏度相当，而LAMP技术的无与伦比的速度使得能够在103 CFUmL 1下获得阳性结果。
The objective of this study was to develop a rapid, sensitive detection assay for the quarantine pathogen Xanthomonas arboricola pv.pruni, causal agent of stone fruit bacterial spot, an economically important disease of Prunus spp. Unique targets were identified from X.arboricola pv.pruni genomes using a comparative genomics pipeline of other Xanthomonas species, subspecies and pathovars, and used to identify specific diagnostic markers. Loop-mediated isothermal amplification (LAMP) was then applied to these markers to provide rapid, sensitive and specific detection. The method developed showed unrivalled specificity with the 79 tested strains and, in contrast to previously established techniques, distinguished between phylogenetically close subspecies such as X.arboricola pv.corylina. The sensitivity of this test is comparable to that of a previously reported TaqMan assay at 103CFUmL1, while the unrivalled speed of LAMP technology enables a positive result to be obtained in