A dual-probe one-pot RPA-CRISPR/Cas12a: a highly sensitive and rapid method for detection of Phytophthora vignae.

IF 3.8 1区农林科学 Q1 AGRONOMY

Pest Management Science Pub Date : 2025-05-26 DOI:10.1002/ps.8932

Mingze Lin,Zhiming Zhou,Zhiting Li,Wanzhen Feng,Chengdong Yang,Zaobing Zhu,Qinghe Chen

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引用次数: 0

Abstract

BACKGROUND Phytophthora vignae is the primary pathogen affecting cowpea plants. Early on-site and accurate detection of the causal pathogen is critical for successful management. Therefore, it is both essential and urgent to create a rapid, sensitive, and accurate detection for P. vignae in cowpeas. RESULTS In this study, the one-pot RPA-CRISPR/Cas12a assay demonstrated the ability to effectively differentiate closely related oomycete species, exhibiting excellent specificity. The entire detection process can be completed within 15-20 min, with results being visually determined using a lateral flow strip (LFS) or by fluorescence detection under blue or ultraviolet (UV) light. Additionally, we have added dual probes to the one-pot RPA-CRISPR/Cas12a assay detection system, this modification improved reaction efficiency and significantly enhanced sensitivity (200 fg μL-1). CONCLUSION This study developed a dual-probe one-pot RPA-CRISPR/Cas12a detection platform that demonstrates rapid and highly sensitive identification of P. vignae. The proposed technology enables field-deployable detection without requiring specialized technical expertise, representing a promising approach with substantial potential for practical applications. © 2025 Society of Chemical Industry.

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双探针一罐RPA-CRISPR/Cas12a：一种高灵敏、快速检测葡萄疫霉菌的方法

背景疫霉（phytophthora vignae）是影响豇豆植株的主要病原体。早期现场和准确检测致病病原体是成功管理的关键。因此，建立一种快速、灵敏、准确的豇豆P. vignae检测方法是十分必要和迫切的。结果在本研究中，单罐RPA-CRISPR/Cas12a实验显示，该方法能够有效区分亲缘关系密切的卵菌种，具有良好的特异性。整个检测过程可在15-20分钟内完成，使用横向流动条（LFS）或蓝色或紫外线（UV）光下的荧光检测可以直观地确定结果。此外，我们在单锅RPA-CRISPR/Cas12a检测系统中添加了双探针，这种修饰提高了反应效率，显著提高了灵敏度（200 fg μL-1）。结论本研究建立了一种双探针单罐RPA-CRISPR/Cas12a检测平台，能够快速、高灵敏度地鉴定葡萄假单胞菌。拟议的技术使现场部署的探测不需要专门的技术知识，代表了一个有前途的方法，具有实际应用的巨大潜力。©2025化学工业协会。

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来源期刊

Pest Management Science 农林科学-昆虫学

CiteScore

7.90

自引率

9.80%

发文量

553

审稿时长

4.8 months

期刊介绍： Pest Management Science is the international journal of research and development in crop protection and pest control. Since its launch in 1970, the journal has become the premier forum for papers on the discovery, application, and impact on the environment of products and strategies designed for pest management. Published for SCI by John Wiley & Sons Ltd.