The Plant Pathology Journal

The Korean Society of Plant Pathology (한국식물병리학회)
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Application of Rapid and Reliable Detection of Cymbidium Mosaic Virus by Reverse Transcription Recombinase Polymerase Amplification Combined with Lateral Flow Immunoassay

  • Do-Hyun, Kim (Department of Agricultural Biology, Jeonbuk National University) ;
  • Rae-Dong, Jeong (Department of Applied Biology, Institute of Environmentally Friendly Agriculture, Chonnam National University) ;
  • Sena, Choi (Horticulture and Herbal Crop Environment Division, National Institute of Horticulture and Herbal Science, Rural Development Administration) ;
  • Ho-Jong, Ju (Department of Agricultural Biology, Jeonbuk National University) ;
  • Ju-Yeon, Yoon (Department of Plant Protection and Quarantine, Jeonbuk National University)
  • Received : 2022.10.30
  • Accepted : 2022.11.15
  • Published : 2022.12.01
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Abstract

Cymbidium mosaic virus (CymMV) is one of economically important viruses that cause significant losses of orchids in the world. In the present study, a reverse transcription recombinase polymerase amplification (RT-RPA) assay combined with a lateral flow immunostrip (LFI) assay was developed for the detection of CymMV in orchid plants. A pair of primers containing fluorescent probes at each terminus that amplifies highly specifically a part of the coat protein gene of CymMV was determined for RT-RPA assay. The RT-RPA assay involved incubation at an isothermal temperature (39℃) and could be performed rapidly within 30 min. In addition, no cross-reactivity was observed to occur with odontoglossum ringspot virus and cymbidium chlorotic mosaic virus. The RT-RPA with LFI assay (RT-RPA-LFI) for CymMV showed 100 times more sensitivity than conventional reverse transcription polymerase chain reaction (RT-PCR). Furthermore, the RT-PCR-LFI assay demonstrated the simplicity and the rapidity of CymMV detection since the assay did not require any equipment, by comparing results with those of conventional RT-PCR. On-site application of the RT-RPA-LFI assay was validated for the detection of CymMV in field-collected orchids, indicating a simple, rapid, sensitive, and reliable method for detecting CymMV in orchids.

Keywords

Acknowledgement

This study was supported by a grant of Cooperative Research Program for Agriculture Science and Technology Development (Project no. PJ01499503), Rural Development Administration, Republic of Korea and supported by "Research Base Construction Fund Support Program" funded by Jeonbuk National University in 2022.

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