A dual-probe one-pot RPA-CRISPR/Cas12a: a highly sensitive and rapid method for detection of Phytophthora vignae
文献类型： 外文期刊
作者： Lin, Mingze 1 ; Zhou, Zhiming 1 ; Li, Zhiting 2 ; Feng, Wanzhen 1 ; Yang, Chengdong 1 ; Zhu, Zaobing 3 ; Chen, Qinghe 1 ;
作者机构： 1.Hainan Univ, Sanya Inst Breeding & Multiplicat, Sch Breeding & Multiplicat, Sanya 572025, Peoples R China
2.Hainan Univ, Sch Trop Agr & Forestry, Key Lab Green Prevent & Control Trop Plant Dis & P, Minist Educ, Haikou, Peoples R China
3.Jiangsu Acad Agr Sci, Inst Plant Protect, Nanjing, Peoples R China
关键词： cowpea; dual-probes; one-pot RPA-CRISPR/Cas12a; visual detection
期刊名称：PEST MANAGEMENT SCIENCE （ 影响因子：3.8； 五年影响因子：4.3 ）
ISSN： 1526-498X
年卷期： 2025 年 81 卷 9 期
页码：
收录情况： SCI
摘要： BACKGROUNDPhytophthora 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.RESULTSIn 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 mu L-1).CONCLUSIONThis 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. (c) 2025 Society of Chemical Industry.
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