Development and application of Rc-RPA-LFD for the rapid detection of Rhizoctonia cerealis.
Development and application of Rc-RPA-LFD for the rapid detection of Rhizoctonia cerealis.
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DOI：
10.13926/j.cnki.apps.000342
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发表时间：
2020-01-01
期刊：
影响因子：
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通讯作者：
Pan, Y. M.
中科院分区：
文献类型：
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作者：
Ju, Yu-Liang;Shen, Peng-Fei;Pan, Y. M.
小麦丝核菌是引起小麦纹枯病的土传植物病原真菌，造成严重的经济损失。本研究建立了重组酶聚合酶扩增结合侧流试纸条技术（Rc-RPA-LFD），用于快速、灵敏地检测R。麦片Rc-RPA-LFD测定可以在38°C的等温温度下在30分钟内在没有PCR热循环仪的情况下完成。根据ITS区设计的RPA引物和探针对红曲霉具有高度的特异性。麦片Rc-RPA-LFD法的检测限为1 pg.穆尔-1，与常规PCR法的检测灵敏度相当。此外，Rc-RPA-LFD检测法可检测R.从田间土壤样品中检测禾谷镰刀菌，与常规PCR检测相比没有显着差异。Rc-RPA-LFD方法的简便、快速和实用性表明，Rc-RPA-LFD方法是一种很有前途的分子诊断方法，可准确、快速地检测R.麦片
Rhizoctonia cerealis is a soil-borne phytopathogenic fungus that causes wheat sharp eyespot, resulting in serious economic losses. In this study, the recombinase polymerase amplification combined with lateral-flow dipstick technology (Rc-RPA-LFD) was developed for the rapid and sensitivity detection of R. cerealis. The Rc-RPA-LFD assay could be completed at isothermal temperature of 38°C within 30 min without PCR thermal cyclers. The RPA primers and probe designed based on ITS region, showed high specificity to R. cerealis. The detection limit of Rc-RPA-LFD assay was 1 pg.muL-1 fungal genomic DNA, showed an equal sensitivity to that of conventional PCR. In addition, the Rc-RPA-LFD assay could detect R. cerealis from field soil samples, showing no significant differences compared to conventional PCR assay. The simplicity, rapidly and practicability all indicated that Rc-RPA-LFD assay will be a promising molecular diagnosis for the accurate and rapid detection of R. cerealis.