The Plant Pathology Journal

The Korean Society of Plant Pathology (한국식물병리학회)
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Development and Application of Reverse Transcription Nanoplate-Based Digital PCR Assay for Sensitive and Accurate Detection of Rice Black-Streaked Dwarf Virus in Cereal Crops

  • Hyo-Jeong Lee (Department of Applied Biology, Chonnam National University) ;
  • Hae-Jun Kim (Department of Applied Biology, Chonnam National University) ;
  • Sang-Min Kim (Crop Foundation Research Division, National Institute of Crop Science, Rural Development Administration) ;
  • Rae-Dong Jeong (Department of Applied Biology, Chonnam National University)
  • Received : 2024.03.05
  • Accepted : 2024.06.25
  • Published : 2024.08.01
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Abstract

The emergence of rice black-streaked dwarf virus (RBSDV) poses a significant threat to global cereal crop cultivation, necessitating the urgent development of reliable detection and quantification techniques. This study introduces a reliable approach for the precise and sensitive quantification of the RBSDV in cereal crop samples, employing a reverse transcription digital polymerase chain reaction (RT-dPCR) assay. We assessed the specificity and sensitivity of the RT-dPCR assay proposed for precise RBSDV detection and quantification. Our findings demonstrate that RT-dPCR was specific for detection of RBSDV, with no cross-reactivity observed with other viruses infecting cereal crops. The RT-dPCR sensitivity was over 10 times that of RT-quantitative PCR (RT-qPCR). The detection limit of RT-dPCR was 0.096 copies/㎕. In addition, evaluation of RT-dPCR assay with field samples was conducted on 60 different cereal crop samples revealed that RT-dPCR (45/60) exhibited superior accuracy compared with RT-qPCR (23/60). In this study, we present a specific and accurate RT-dPCR assay for the detection and quantification of RBSDV.

Keywords

Acknowledgement

This work was carried out with the support of Cooperative Research Program for Agriculture Science and Technology Development (Project No. PJ014995022023) Rural Development Administration, Republic of Korea.

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