Abstract
Maize chlorotic mottle virus (MCMV), an important quarantine virus, causes lethal necrosis in maize when coinfected with a potyvirid, which is seriously threatening the production of maize worldwide. In this study, recombinase polymerase amplification (RPA), a novel isothermal DNA amplification and detection technique, was developed to detect MCMV in maize crops. A pair of specific primers was designed based on the conserved sequences of the MCMV coat protein region. The RT-RPA assay was carried out as an isothermal reaction at 38 °C that was complete within 30 min, and no cross-reactivity was detected with other viruses infecting maize in China. The limit of detection of the RT-RPA assay was tenfold lower than that of ordinary RT-PCR. Moreover, this method was successfully applied to test field-collected samples. The newly developed RT-RPA assay offers a reliable, sensitive and efficient method for rapid detection of MCMV in maize in equipment-limited diagnostic laboratories and on-site facilities.
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Acknowledgements
We thank Prof. Zaifeng Fan (China Agricultural University) for providing MCMV, SCMV, RBSDV and PenMV. This research was funded by grants from the Natural Science Foundation of China (31801702).
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ZX and YW conceived and designed the experiments. JJ and YJ performed the experiments. JJ and YJ wrote the paper. ZX, MA and YW edited the paper.
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Jiao, Y., Jiang, J., An, M. et al. Recombinase polymerase amplification assay for rapid detection of maize chlorotic mottle virus in maize. Arch Virol 164, 2581–2584 (2019). https://doi.org/10.1007/s00705-019-04361-3
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DOI: https://doi.org/10.1007/s00705-019-04361-3