Rapid and quantitative detection of citrus huanglongbing bacterium 'Candidatus Liberibacter asiaticus' by real-time fluorescent loop-mediated isothermal amplification assay in China

文献类型： 外文期刊

第一作者： Wu, Xuhui

作者： Wu, Xuhui;Meng, Chunliang;Wang, Guihua;Liu, Yuanzheng;Zhang, Xin;Yi, Kexian;Peng, Jun

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关键词： Huanglongbing (HLB);Candidatus Liberibacter asiaticus (Las);Quantitative detection;Visual inspection;Real-time fluorescence loop-mediated isothermal amplification (RealAmp)

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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年卷期：

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收录情况： SCI

摘要： Huanglongbing (HLB) is a devastating citrus disease that is associated with bacteria of the species 'Candidatus Liberibacter asiaticus (Las)' in China. All currently popular cultivars are susceptible to HLB, and there are no effective approaches to control the disease once the plants have been infected. Therefore, rapid and easy diagnostic tools are needed for maintaining clean nursery stock and inoculum removal strategies to control HLB. A real-time fluorescence loop-mediated isothermal amplification (RealAmp) targeting16S rDNA was developed for the rapid and quantitative detection of Las in China. The detection sensitivity of the RealAmp assay was approximately 1 pg/mu l template DNA (equal to 2.4 x 10(5) target DNA copies/mu l) and no cross-reaction was observed with other pathogens or virus-free seedlings. Besides the quantitative detection using SYTO-9 fluorescent dye, an improved closed-tube visual inspection technique using SYBR Green I staining was developed for rapid screening of samples and minimize the risk of cross-contamination. The RealAmp assay is an alternative quantitative method, which would be used as a routine detection service for nursery materials in China. (C) 2016 Elsevier Ltd. All rights reserved.

分类号： S432.1

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