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首页> 外文期刊>Plant Pathology >Genomics-informed design of loop-mediated isothermal amplification for detection of phytopathogenic Xanthomonas arboricola pv. pruni at the intraspecific level
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Genomics-informed design of loop-mediated isothermal amplification for detection of phytopathogenic Xanthomonas arboricola pv. pruni at the intraspecific level

机译：基因组学设计的环介导等温扩增技术，用于检测植物致病性黄单胞菌。种内水平的pruni

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摘要

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 10~3 CFU rnL~(-1), while the unrivalled speed of LAMP technology enables a positive result to be obtained in <15 min. The developed assay can be used with real-time fluorescent detectors for quantitative results as well as with DNA-staining dyes to function as a simplified strategy for on-site pathogen detection.
机译：这项研究的目的是开发一种快速，灵敏的检疫病原体Xanthomonas arboricola pv检测方法。 pruni，核果细菌斑的病原体，一种经济上重要的李子病。从X. arboricola pv确定了唯一的目标。 pruni基因组使用其他Xanthomonas物种，亚种和Pathovars的比较基因组学方法，用于鉴定特定的诊断标记。然后将环介导的等温扩增（LAMP）应用于这些标记，以提供快速，灵敏和特异性的检测。所开发的方法对79种测试菌株显示出无与伦比的特异性，并且与先前建立的技术相比，在系统发育上接近的亚种（如X. arboricola pv）之间进行了区分。 corylina。该测试的灵敏度与以前报道的TaqMan测定法在10〜3 CFU rnL〜（-1）时的灵敏度相当，而LAMP技术的无与伦比的速度使得在<15分钟内即可获得阳性结果。所开发的测定法可以与实时荧光检测器一起使用以定量结果，也可以与DNA染色染料一起使用，作为现场病原体检测的简化策略。

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