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Pest Management Science
Article . 2024 . Peer-reviewed
License: CC BY NC ND
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Article . 2025
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Rapid and equipment‐free identification of papaya mealybug Paracoccus marginatus based on RPA‐CRISPR/Cas12a

Authors: Chen, Yan‐ting; Shi, Meng‐zhu; Chen, Yan; Zhao, Jian‐wei; Yang, Xiu‐juan; Fu, Jian‐wei; Desneux, Nicolas; +1 Authors

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

Abstract

AbstractBACKGROUNDParacoccus 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.RESULTSA 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.CONCLUSIONThis 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.

Country
France
Keywords

Carica, CRISPR/Cas12a, Paracoccus, fluorescence visualization, Recombinases, papaya mealybug, lateral flow strip, [SDV.BA.ZI] Life Sciences [q-bio]/Animal biology/Invertebrate Zoology, Animals, CRISPR-Cas Systems, recombinase polymerase amplification (RPA), Nucleic Acid Amplification Techniques, Research Article

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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
4
Top 10%
Average
Top 10%
Green
hybrid
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