Rapid and equipment-free identification of papaya mealybug Paracoccus marginatus based on RPA-CRISPR/Cas12a

文献类型： 外文期刊

第一作者： Chen, Yan-Ting

作者： Chen, Yan-Ting;Shi, Meng-Zhu;Chen, Yan;Zhao, Jian-Wei;Yang, Xiu-Juan;Li, Jian-Yu;Shi, Meng-Zhu;Fu, Jian-Wei;Shi, Meng-Zhu;Desneux, Nicolas;Li, Jian-Yu

作者机构：

关键词： papaya mealybug; recombinase polymerase amplification (RPA); CRISPR/Cas12a; fluorescence visualization; lateral flow strip

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 1 期

页码：

收录情况： SCI

摘要： BACKGROUND: 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. RESULTS: 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. CONCLUSION: 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. (c) 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

分类号：

  • 相关文献
作者其他论文 更多>>

微信小程序