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A multiplex RT-PCR method to detect papaya meleira virus complex in adult pre-flowering plants

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Abstract

Papaya sticky disease (PSD), which can destroy orchards, was first attributed to papaya meleira virus (PMeV). However, the discovery of papaya meleira virus 2 (PMeV2) associated with PSD plants impose the need to detect this viral complex. We developed a multiplex RT-PCR (mPCR) technique capable of detecting two viruses in a single assay from pre-flowering plant samples, which is a useful tool for early diagnosis of PSD. We also determined the limit of detection (LOD) using asymmetric plasmid dilutions of both PMeV and PMeV2, which revealed that a higher titer of one virus prevents detection of the other. Thus, this technique is an alternative method for detecting PMeV and PMeV2 in a single reaction.

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Abbreviations

PSD:

papaya sticky disease

PMeV:

papaya meleira virus

PMeV2:

papaya meleira virus 2

mPCR:

multiplex PCR

LOD:

limit of detection

RdRp:

RNA-dependent RNA polymerase

ORF:

open reading frame

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Acknowledgments

T.F. Sá Antunes and S. A. Oliveira would like to thank the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, CAPES (Ministry of Education of Brazil) and M. Maurastoni the Fundação de Amparo à Pesquisa do Espírito Santo, FAPES for a fellowship. P.M.B. Fernandes and José A. Ventura were granted the research productivity award from the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Grant #304719/2014-5 and #308631/2016-1, respectively.

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Authors and Affiliations

  1. Núcleo de Biotecnologia, Universidade Federal do Espírito Santo, Vitória, Espírito Santo, Brazil

    Marlonni Maurastoni, Tathiana F. Sá-Antunes, Scarlett A. Oliveira, Alexandre M. C. Santos, José A. Ventura & Patricia M. B. Fernandes

  2. Instituto Capixaba de Pesquisa, Assistência Técnica e Extensão Rural, Vitória, Epírito Santo, Brazil

    José A. Ventura

Authors
  1. Marlonni Maurastoni
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  2. Tathiana F. Sá-Antunes
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  3. Scarlett A. Oliveira
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  4. Alexandre M. C. Santos
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  5. José A. Ventura
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  6. Patricia M. B. Fernandes
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Correspondence to Patricia M. B. Fernandes.

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The authors declare that they have no conflict of interest.

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Maurastoni, M., Sá-Antunes, T.F., Oliveira, S.A. et al. A multiplex RT-PCR method to detect papaya meleira virus complex in adult pre-flowering plants. Arch Virol 165, 1211–1214 (2020). https://doi.org/10.1007/s00705-020-04588-5

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  • DOI: https://doi.org/10.1007/s00705-020-04588-5

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