Development of a Recombinase Polymerase Amplification and CRISPR-Cas12a-Based Assay for Rapid Detection of Rice Bakanae Disease Caused by Fusarium fujikuroi

文献类型： 外文期刊

第一作者： Li, Hongyu

作者： Li, Hongyu;Hu, Bilian;Li, Hongyu;Cheng, Can;Zhou, Jihua;Niu, Fuan;Sun, Bin;Cao, Liming;Chu, Huangwei;Qiu, Yue;Zhang, Anpeng;Hu, Yingxiong;Dai, Yuting;Xie, Kaizhen;Feng, Zhizun;Ding, Xiaorui;Hu, Bilian;Zhang, Xueqing;Qiu, Yue;Ding, Xiaorui;Zhang, Xueqing

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关键词： CRISPR-Cas12a; RPA; Fusarium fujikuroi; rice bakanae disease; detection

期刊名称：AGRONOMY-BASEL （ 影响因子：3.4； 五年影响因子：3.8 ）

ISSN：

年卷期： 2025 年 15 卷 3 期

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收录情况： SCI

摘要： Fusarium fujikuroi is the primary causal agent of rice bakanae disease, which can lead to substantial yield losses. Developing a rapid, highly specific, and accurate method for detecting F. fujikuroi is crucial for effective surveillance, prevention, and control of rice bakanae disease. In this study, a novel detection assay, RPA-Cas12a-F, was developed by integrating recombinase polymerase amplification (RPA) and Cas12a for the detection of F. fujikuroi. This assay demonstrated a limit of detection (LOD) of 1 copy/mu L of reference plasmid or 0.1 fg/mu L of F. fujikuroi genomic DNA (gDNA). Furthermore, to enable on-site detection, the RPA-Cas12a technique was combined with a lateral flow strip (LFS) for visual readout, thereby developing the RPA-Cas12a-LFS assay. The LOD of the RPA-Cas12a-LFS assay was 1000 copies/mu L of plasmid or 10 fg/mu L of F. fujikuroi gDNA. The RPA-Cas12a-based assays developed in this study enable rapid, highly accurate, sensitive, and specific detection of F. fujikuroi, making them a promising tool for on-site detection without the need for expensive equipment and time-consuming methodologies.

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