Detection of four regulated grapevine viruses in a qualitative, single tube real-time PCR with melting curve analysis

M. Aloisio, M. Morelli, V. Elicio, P. Saldarelli, B. Ura, B. Bortot, G. M. Severini, A. Minafra

Research output: Contribution to journalArticle

Abstract

The detection of the four grapevine viruses (GLRaV-1, GLRaV-3, GFLV and ArMV) regulated in European Union plant material certification, requires sensitive and specific diagnostic tools. A strategy of simultaneous detection in a real-time single tube amplification was developed, based on the EvaGreen binding dye. The melting curve analysis (MCA) of the amplicons allows a qualitative detection of the four different virus targets in multiplex analysis. A plasmid dilution assay calculated an analytical sensitivity with an amplification threshold up to 100 copies of the target sequences. A small cohort of field grapevine samples, with a known status of infection by mixtures of the target viruses or free of them, respectively, was successfully tested for the evaluation of the amplicons Tm.

Original languageEnglish
Pages (from-to)42-47
Number of pages6
JournalJournal of Virological Methods
Volume257
DOIs
Publication statusPublished - Jul 1 2018

Keywords

  • EvaGreen
  • Grapevine
  • Melting curve
  • Multiplex Real-time PCR
  • Viruses

ASJC Scopus subject areas

  • Virology

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