The Plant Pathology Journal

The Korean Society of Plant Pathology (한국식물병리학회)
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Development of Recombinase Polymerase Amplification Combined with Lateral Flow Strips for Rapid Detection of Cowpea Mild Mottle Virus

  • Xinyang Wu (College of Life Sciences, China Jiliang University) ;
  • Shuting Chen (College of Life Sciences, China Jiliang University) ;
  • Zixin Zhang (College of Life Sciences, China Jiliang University) ;
  • Yihan Zhang (College of Life Sciences, China Jiliang University) ;
  • Pingmei Li (College of Life Sciences, China Jiliang University) ;
  • Xinyi Chen (College of Life Sciences, China Jiliang University) ;
  • Miaomiao Liu (College of Life Sciences, China Jiliang University) ;
  • Qian Lu (College of Life Sciences, China Jiliang University) ;
  • Zhongyi Li (College of Life Sciences, China Jiliang University) ;
  • Zhongyan Wei (State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, Key Laboratory of Biotechnology in Plant Protection of Ministry of Agriculture and Zhejiang Province, Institute of Plant Virology, Ningbo University) ;
  • Pei Xu (College of Life Sciences, China Jiliang University)
  • Received : 2023.03.02
  • Accepted : 2023.09.10
  • Published : 2023.10.01
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Abstract

Cowpea mild mottle virus (CPMMV) is a global plant virus that poses a threat to the production and quality of legume crops. Early and accurate diagnosis is essential for effective managing CPMMV outbreaks. With the advancement in isothermal recombinase polymerase amplification and lateral flow strips technologies, more rapid and sensitive methods have become available for detecting this pathogen. In this study, we have developed a reverse transcription recombinase polymerase amplification combined with lateral flow strips (RT-RPA-LFS) method for the detection of CPMMV, specifically targeting the CPMMV coat protein (CP) gene. The RT-RPA-LFS assay only requires 20 min at 40℃ and demonstrates high specificity. Its detection limit was 10 copies/µl, which is approximately up to 100 times more sensitive than RT-PCR on agarose gel electrophoresis. The developed RT-RPA-LFS method offers a rapid, convenient, and sensitive approach for field detection of CPMMV, which contribute to controlling the spread of the virus.

Keywords

Acknowledgement

This work is supported by National Nature Science Foundation of China (32202470), Key Research Program of Zhejiang Province (2021C02041), and State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products (2021DC700024-KF202217).

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