A real-time recombinase polymerase amplification assay for fast and accurate detection of Ditylenchus destructor

文献类型： 外文期刊

第一作者： Gao, Bo

作者： Gao, Bo;Ma, Juan;Li, Xiuhua;Wang, Rongyan;Chen, Shulong

作者机构：

关键词： Ditylenchus destructor; Plant-parasitic nematodes; Real-time; Recombinase polymerase amplification; Detection

期刊名称：MOLECULAR AND CELLULAR PROBES （ 影响因子：3.285； 五年影响因子：2.781 ）

ISSN： 0890-8508

年卷期： 2022 年 61 卷

页码：

收录情况： SCI

摘要： Ditylenchus destructor is a plant-parasitic nematode that seriously infests sweet potato crop in China. Thus, fast and accurate detection of D. destructor in soil and plant tissue samples is of great significance. In this study, a real-time recombinase polymerase amplification (RPA) assay was developed for the rapid and accurate detection of D. destructor in various samples. The RPA assay could be easily operated and detected as low as 1/500 individual J4 nematode DNA per reaction in 20 min at 39 degrees C with high specificity. The assay meets the requirements of rapid detection prior to port quarantine as well as on-site real-time detection and can be applied to detect the parasite in soil and plant samples. The modified gDNA extraction method for a single nematode established in this study significantly reduced the time of detection and improved the applicability of the real-time RPA assay for on-site detection in different environments. The real-time RPA assay to detect D. destructor will be useful for epidemiological investigations in the field as well as for quarantine processes in the sweet potato and potato trade.

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