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Simultaneous detection of Cymbidium mosaic virus and Odontoglossum ringspot virus in orchids using multiplex RT-PCR

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Abstract

A system for simultaneous detection of two orchid-infecting viruses was developed and applied to several orchid species. The detection system involved multiplex reverse transcription-polymerase chain reaction (RT-PCR) and could simultaneously identify Cymbidium mosaic virus (CymMV) and Odontoglossum ringspot virus (ORSV) from the orchid species studied. Multiplex RT-PCR was conducted using two virus-specific primer pairs and an internal control pair of primers to amplify the CymMV and ORSV coat protein regions, and orchid 18S rDNA, respectively. For optimization of multiplex RT-PCR conditions, serial dilutions of total RNA and cDNA were performed and the detection limit of the system was evaluated. The optimized multiplex detection system for CymMV and ORSV was applied to various orchid species, including several cultivars of Doritaenopsis, Cymbidium, Dendrobium, and Phalaenopsis to test the efficacy of this method. Our results indicate that the multiplex RT-PCR detection system will be a rapid, simple, and precise diagnosis tool in a range of orchid species.

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References

  1. F.W. Zettler, N.J. Ko, G.C. Wisler, M.S. Elliott, S.M. Wong, Plant Dis. 74, 621–626 (1990)

    Article  Google Scholar 

  2. K.W. Koh, H. Lu, M. Chan, Plant Sci. 228, 26–38 (2014)

    Article  CAS  PubMed  Google Scholar 

  3. S.M. Wong, C.G. Chng, Y.H. Lee, K. Tan, F.W. Zettler, Crop Protect. 13, 235–239 (1994)

    Article  Google Scholar 

  4. A.J. Eun, M.L. Seoh, S.M. Wong, J. Virol. Methods 87, 151–160 (2000)

    Article  CAS  PubMed  Google Scholar 

  5. M.L. Seoh, S.M. Wong, L. Zhang, J. Virol. Methods 72, 197–204 (1998)

    Article  CAS  PubMed  Google Scholar 

  6. S.C. Lee, Y.C. Chang, Plant Pathol. Bull. 15, 187–196 (2006)

    CAS  Google Scholar 

  7. M.S. Lee, M.J. Yang, Y.C. Hseu, G.H. Lai, W.T. Chang, Y.H. Hsu, M.K. Lin, J. Virol. Methods 173, 43–48 (2011)

    Article  CAS  PubMed  Google Scholar 

  8. H.Y. Lin, C.H. Huang, S.H. Lu, I.T. Kuo, L.K. Chau, Biosens. Bioelectron. 51, 371–378 (2014)

    Article  CAS  PubMed  Google Scholar 

  9. K. Yamane, K. Oyama, E. Iuchi, H. Ogawa, T. Suzuki, T. Natsuaki, J. Phytopathol. 156, 268–373 (2008)

    Article  CAS  Google Scholar 

  10. S.H. Choi, J.S. Hong, K.H. Ryu, Korean J. Hort. Sci. Tech. 28, 103–107 (2010)

    CAS  Google Scholar 

  11. H. Uga, S. Tsuda, Phytopathology 95, 166–171 (2005)

    Article  CAS  PubMed  Google Scholar 

  12. A. Roy, A. Fayad, G. Barthe, R.H. Brlansky, J. Virol. Methods 129, 47–55 (2005)

    Article  CAS  PubMed  Google Scholar 

  13. W. Menzel, W. Jelkmann, E. Maiss, J. Virol. Methods 99, 81–92 (2002)

    Article  CAS  PubMed  Google Scholar 

  14. P.A. Ajjikuttira, C.L. Lom-Ho, M.H. Woon, K.H. Ryu, C.A. Chang, C.S. Loh, S.M. Wong, Arch. Virol. 147, 1943–1954 (2002)

    Article  CAS  PubMed  Google Scholar 

  15. J.Y. Yoon, B.N. Chung, G.S. Choi, S.K. Choi, Virus Genes 44, 323–328 (2012)

    Article  CAS  PubMed  Google Scholar 

  16. S.Y. Park, H.N. Murthy, K.Y. Paek, Plant Sci. 164, 919–923 (2003)

    Article  CAS  Google Scholar 

  17. G.C. Wey, T.Y. Shyuu, W.H. Chen, Taiwan Sugar 48, 22–29 (2001)

    Google Scholar 

  18. S.P. Vaughan, M. Grisoni, M.H. Kumagai, A.R. Kuehnle, Arch. Virol. 153, 1185–1189 (2008)

    Article  CAS  PubMed  Google Scholar 

  19. Y.X. Zheng, B.N. Shen, C.C. Chen, F.J. Jan, Eur. J. Plant Pathol. 128, 1–5 (2010)

    Article  Google Scholar 

  20. N.A. Raymond, A.L. Dann, P.A. Cross, C.R. Wilson, Arch. Virol. 159, 3095–3099 (2014)

    Article  Google Scholar 

Download references

Acknowledgments

This work was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (2014R1A1A2056541).

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Authors and Affiliations

  1. Plant Virus Genbank, Department of Horticulture, Biotechnology and Landscape Architecture, Seoul Women’s University, 621 Hwarangro, Nowon, Seoul, 139-774, Korea

    Su Min Kim & Sun Hee Choi

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  1. Su Min Kim
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  2. Sun Hee Choi
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Correspondence to Sun Hee Choi.

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Edited by Thomas Hohn.

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Kim, S.M., Choi, S.H. Simultaneous detection of Cymbidium mosaic virus and Odontoglossum ringspot virus in orchids using multiplex RT-PCR. Virus Genes 51, 417–422 (2015). https://doi.org/10.1007/s11262-015-1258-x

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  • DOI: https://doi.org/10.1007/s11262-015-1258-x

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