Comparison of sensitivity and rapidness of PCR, recombinase polymerase amplification, and RNA-specific amplification for detection of Rice yellow mottle virus

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  • Juma Kevin Maafu
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University
  • Kojima Kenji
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University
  • Takita Teisuke
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University
  • T. Natsuaki Keiko
    Department of International Agricultural Development, Tokyo University of Agriculture
  • Yasukawa Kiyoshi
    Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University

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Abstract

Rice yellow mottle virus (RYMV) is a plant pathogenic virus that often causes a serious damage　to rice production in Africa. In this study, we developed detection systems of RYMV DNA or RNA　using each of PCR, recombinase polymerase amplification (RPA), and an RNA-specific　amplification. The sensitivities were in the range of several　copies of the target DNA for PCR and　RPA and dozens copies of the target RNA for RNA-specific amplification.　The cycle numbers or　reaction times required for amplification from 109 copies of the target DNA or RNA were 15 cycles　(27 min) for the PCR-based system and 5 min for RPA- or RNA-specific amplification-based　systems.　These results suggested that isothermal RPA and RNA-specific amplification-based　detection systems of RYMV will be more suitable for quick detection of RYMV-infected rice plants　than the PCR-based one.

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