Chen Yan-Ting, Shi Meng-Zhu, Chen Yan, Zhao Jian-Wei, Yang Xiu-Juan, Fu Jian-Wei, Desneux Nicolas, Li Jian-Yu
Fujian Key Laboratory for Monitoring and Integrated Management of Crop Pests, Institute of Plant Protection, Fujian Academy of Agricultural Sciences, Fuzhou, China.
Institute of Quality Standards and Testing Technology for Agro-Products, Fujian Academy of Agricultural Sciences, Fuzhou, China.
Pest Manag Sci. 2025 Jan;81(1):230-239. doi: 10.1002/ps.8425. Epub 2024 Sep 25.
Paracoccus marginatus has invaded many countries, spreading rapidly and causing significant economic losses to crops. Accurate detection during the monitoring process is critical to prevent its expansion into new areas, therefore it is necessary to develop efficient and reliable detection methods. Traditional detection methods are time-consuming and instrument-dependent owing to the morphological similarities and small sizes of P. marginatus and other mealybugs, therefore establishing an efficient, rapid, and sensitive method for field detection in resource-limited settings is critical.
A sensitive and rapid detection system was developed to detect P. marginatus using recombinase polymerase amplification (RPA) combined with clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a. The RPA-CRISPR/Cas12a assay distinguished P. marginatus from 10 other mealybugs. The entire process can be completed in approximately an hour, and the identification results can be determined by the naked eye using lateral flow strips or a portable mini-UV torch. A method was developed to extract DNA from P. marginatus within 5 min. This method was combined with the RPA-CRISPR/Cas12a assay to achieve rapid and simple detection. In addition, two portable thermos cups with temperature displays were used to maintain the reagents and assay reactions in the field.
This assay represents the first application of portable and easily available items (mini-UV torch and thermos cup) based on the combination of RPA and CRISPR/Cas12a for rapid pest detection. This method is rapid, highly specific, and instrument-flexible, allowing for the early monitoring of P. marginatus in the field. This study provides guidance for the development of suitable management strategies. © 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.
边缘新圆蚧已入侵许多国家，传播迅速，给农作物造成重大经济损失。在监测过程中进行准确检测对于防止其扩散到新区域至关重要，因此有必要开发高效可靠的检测方法。由于边缘新圆蚧与其他粉蚧在形态上相似且体型较小，传统检测方法既耗时又依赖仪器，因此在资源有限的环境中建立一种高效、快速且灵敏的现场检测方法至关重要。
开发了一种灵敏且快速的检测系统，利用重组酶聚合酶扩增（RPA）结合成簇规律间隔短回文重复序列（CRISPR）/Cas12a来检测边缘新圆蚧。RPA-CRISPR/Cas12a检测法可将边缘新圆蚧与其他10种粉蚧区分开来。整个过程大约可在一小时内完成，鉴定结果可使用侧向流动试纸条或便携式微型紫外线手电筒通过肉眼确定。开发了一种在5分钟内从边缘新圆蚧中提取DNA的方法。该方法与RPA-CRISPR/Cas12a检测法相结合，实现了快速简便的检测。此外，还使用了两个带有温度显示的便携式保温杯在现场维持试剂和检测反应。
该检测法代表了基于RPA和CRISPR/Cas12a相结合的便携式且易于获取的物品（微型紫外线手电筒和保温杯）在害虫快速检测中的首次应用。该方法快速、特异性高且仪器灵活性强，能够在田间对边缘新圆蚧进行早期监测。本研究为制定合适的管理策略提供了指导。© 2024作者。由John Wiley & Sons Ltd代表化学工业协会出版的《害虫管理科学》。
