Development of a Loop-Mediated Isothermal Amplification Method for the Rapid Detection of Phytopythium vexans

文献类型： 外文期刊

第一作者： Wang, Tuhong

作者： Wang, Tuhong;Ji, Haojun;Yu, Yongting;Cheng, Yi;Li, Zhimin;Chen, Jia;Guo, Litao;Xu, Jianping;Gao, Chunsheng;Wang, Tuhong;Ji, Haojun;Yu, Yongting;Cheng, Yi;Li, Zhimin;Chen, Jia;Guo, Litao;Xu, Jianping;Gao, Chunsheng;Ji, Haojun;Wang, Xiaojie;Xu, Jianping

作者机构：

关键词： Phytopythium vexans; loop mediated isothermal amplification; internal transcribed spacer (ITS) sequence; hydroxynaphthol blue (HNB); diagnosis

期刊名称：FRONTIERS IN MICROBIOLOGY （ 影响因子：5.64； 五年影响因子：6.32 ）

ISSN：

年卷期： 2021 年 12 卷

页码：

收录情况： SCI

摘要： Brown root rot caused by Phytopythium vexans is a new destructive root disease on many plants such as Gingko, Citrus, kiwifruit, and ramie. The establishment of loop-mediated isothermal amplification (LAMP) technology for detecting P. vexans can help monitor and control brown root rot quickly, efficiently, and accurately. LAMP technology is known for its simplicity, sensitivity, and speed; and it does not require any specialized equipment - a water bath or a thermoblock is sufficient for isothermal amplifications. LAMP products can be visualized by using hydroxy naphthol blue (HNB) dye or agarose gel electrophoresis. In this study, by searching and comparing the internal transcribed spacer (ITS) sequences of P. vexans and the related species in oomycete genera Pythium, Phytopythium, and Phytophthora, we designed specific primers targeting the ITS gene region of P. vexans. Using HNB dye, we established a LAMP technique for rapid detection of P. vexans by visible color change. In addition, we optimized the protocol to enhance both sensitivity and specificity for P. vexans detection. Under the optimized condition, our protocol based on LAMP technology could detect as low as 24 copies of the P. vexans genomic DNA, which is similar to 100 times more sensitive than conventional PCR. This method can successfully detect P. vexans using cell suspensions from P. vexans - infected ramie root tissues.

分类号：

  • 相关文献
作者其他论文 更多>>

微信小程序