Rapid and Visual Detection of Pyricularia oryzae Using Coupled Recombinase Polymerase Amplification-Lateral Flow Dipstick Assay

文献类型： 外文期刊

第一作者： Du, Yan

作者： Du, Yan;Qi, Zhongqiang;Yu, Junjie;Zhang, Rongsheng;Yu, Mina;Cao, Huijuan;Song, Tianqiao;Pan, Xiayan;Wang, Shuchen;Liu, Yongfeng;Yang, Jun;Xiong, Qin;Liu, Yongfeng

作者机构：

关键词： disease diagnosis; lateral flow dipstick; Pyricularia oryzae; recombinase polymerase amplification

期刊名称：PLANT DISEASE （ 影响因子：4.4； 五年影响因子：4.8 ）

ISSN： 0191-2917

年卷期： 2025 年 109 卷 5 期

页码：

收录情况： SCI

摘要： Rice blast, caused by Pyricularia oryzae, is one of the most destructive fungal diseases in rice, severely impacting rice production worldwide every year. Rapid, accurate, and visual detection of P. oryzae is essential for more effective prevention and control. In this study, we developed a recombinase polymerase amplification-lateral flow dipstick (RPA-LFD) assay to detect P. oryzae. Species-specific RPA primer pairs and probe were designed based on the target gene MGG_15975. The optimized reaction temperature and time were set at 37 degrees C and 25 min, respectively. Specificity analysis showed that the assay could specifically detect P. oryzae isolates from rice, whereas other fungal species or Pyricularia species from grasses were not detected. Additionally, this assay demonstrated high sensitivity, capable of detecting as low as 10-2 ng/mu l of P. oryzae genomic DNA, which was found to be 100 times more sensitive than conventional PCR. Furthermore, using this assay, P. oryzae was effectively detected in diseased leaves in rice fields and could also be identified at an early stage of infection before obvious lesions appeared in artificially inoculated rice seedlings. Therefore, the RPA-LFD assay developed in our study for the detection of P. oryzae is rapid, highly sensitive, and efficient, which has the potential application for early diagnosis of P. oryzae infection in rice fields.

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