Generic RT-PCR tests for detection and identification of tospoviruses

A. Hassani-Mehraban, M. Westenberg, J.T.J. Verhoeven, B.T.L.H. van de Vossenberg, R. Kormelink, J.W. Roenhorst

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Abstract

A set of tests for generic detection and identification of tospoviruses has been developed. Based on a multiple sequence alignment of the nucleocapsid gene and its 5' upstream untranslated region sequence from 28 different species, primers were designed for RT-PCR detection of tospoviruses from all recognized clades, i.e. the American, Asian and Eurasian clades, and from the small group of distinct and floating species. Pilot experiments on isolates from twenty different species showed that the designed primer sets successfully detected all species by RT-PCR, as confirmed by nucleotide sequence analysis of the amplicons. In a final optimized design, the primers were applied in a setting of five RT-PCR tests. Seven different tospoviruses were successfully identified from diagnostic samples and in addition a non-described tospovirus species from alstroemeria plants. The results demonstrate that the newly developed generic RT-PCR tests provide a relevant tool for broad detection and identification of tospoviruses in plant quarantine and diagnostic laboratories.

Original languageEnglish
Pages (from-to)89-96
JournalJournal of Virological Methods
Volume233
DOIs
Publication statusPublished - 2016

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Keywords

  • Bunyaviridae
  • Clade-specific primers
  • Diagnostics
  • Nucleocapsid gene
  • Plant virus

Cite this

Hassani-Mehraban, A., Westenberg, M., Verhoeven, J. T. J., van de Vossenberg, B. T. L. H., Kormelink, R., & Roenhorst, J. W. (2016). Generic RT-PCR tests for detection and identification of tospoviruses. Journal of Virological Methods, 233, 89-96. https://doi.org/10.1016/j.jviromet.2016.03.015