Phurbu Dorji, Feng Zhipeng, Cai Lei, Liu Fang
Xizang Key Laboratory of Plateau Fungi, Institute of Plateau Biology of Xizang Autonomous Region, Lhasa, China Institute of Plateau Biology of Xizang Autonomous Region Lhasa China.
State Key Laboratory of Microbial Diversity and Innovative Utilization, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China Chinese Academy of Sciences Beijing China.
IMA Fungus. 2025 Jun 10;16:e153604. doi: 10.3897/imafungus.16.153604. eCollection 2025.
With the increase in cross-border transmission in the context of globalization, the necessity for developing rapid and accurate detection methods for plant pathogens has become critical. This study introduces a recombinase polymerase amplification (RPA) technique combined with CRISPR/Cas12a cleavage and fluorescence-based detection systems (FRB) or paper-based lateral flow strips (PLFS) for the rapid on-site detection of invasive alien fungi, specifically and , which pose significant threats to agriculture and biodiversity. The results demonstrate that either RPA-CRISPR/Cas12a-FRB or RPA-CRISPR/Cas12a-PLFS can accurately detect the target species within 30 min, with a sensitivity of up to 10 pg/μL. These portable and easy-to-use assays are suitable for rapid on-site screening of plant pathogenic fungi in plant tissues, enabling applications in disease control and port quarantine.
随着全球化背景下跨境传播的增加，开发快速、准确的植物病原体检测方法变得至关重要。本研究介绍了一种重组酶聚合酶扩增（RPA）技术，该技术结合了CRISPR/Cas12a切割和基于荧光的检测系统（FRB）或基于试纸条的侧向流动条（PLFS），用于快速现场检测外来入侵真菌，特别是对农业和生物多样性构成重大威胁的 和 。结果表明，RPA-CRISPR/Cas12a-FRB或RPA-CRISPR/Cas12a-PLFS均可在30分钟内准确检测到目标物种，灵敏度高达10 pg/μL。这些便携式且易于使用的检测方法适用于对植物组织中的植物病原真菌进行快速现场筛查，可应用于疾病控制和口岸检疫。
