Lin Mingze, Zhou Zhiming, Li Zhiting, Feng Wanzhen, Yang Chengdong, Zhu Zaobing, Chen Qinghe
School of Breeding and Multiplication, Sanya Institute of Breeding and Multiplication, Hainan University, Sanya, China.
Key Laboratory of Green Prevention and Control of Tropical Plant Diseases and Pests, Ministry of Education, School of Tropical Agriculture and Forestry, Hainan University, Haikou, China.
Pest Manag Sci. 2025 Sep;81(9):5784-5795. doi: 10.1002/ps.8932. Epub 2025 May 26.
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.
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).
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.
豇豆疫霉是影响豇豆植株的主要病原菌。早期进行现场准确检测致病病原菌对于成功防治至关重要。因此，创建一种针对豇豆中豇豆疫霉的快速、灵敏且准确的检测方法既至关重要又迫在眉睫。
在本研究中，一锅法重组酶聚合酶扩增（RPA）-CRISPR/Cas12a检测方法展现出有效区分密切相关卵菌物种的能力，具有出色的特异性。整个检测过程可在15 - 20分钟内完成，结果可通过侧向流动试纸条（LFS）进行肉眼判定，或在蓝色或紫外（UV）光下通过荧光检测来确定。此外，我们在一锅法RPA-CRISPR/Cas12a检测系统中添加了双探针，这种改进提高了反应效率并显著增强了灵敏度（200 fg μL）。
本研究开发了一种双探针一锅法RPA-CRISPR/Cas12a检测平台，该平台可快速且高度灵敏地鉴定豇豆疫霉。所提出的技术无需专业技术知识即可用于现场检测，是一种具有巨大实际应用潜力的有前景的方法。© 2025化学工业协会。
