Rapid and specific detection of wheat spindle streak mosaic virus using RT-LAMP in durum wheat crude leaf extract

PLoS One. 2024 Feb 29;19(2):e0299078. doi: 10.1371/journal.pone.0299078. eCollection 2024.

Abstract

To accurately determine the spread of any pathogen, including plant viruses, a quick, sensitive, cost-effective, point-of-care diagnostic assay is necessary. Wheat spindle streak mosaic virus (WSSMV) is a Bymovirus, transmitted by the plasmodiophorid Polymyxa graminis Led, which causes yellow mosaic and reduces the grain yield in wheat. Currently, detection protocols for WSSMV use ELISA or more sensitive PCR-based approaches requiring specialized laboratory and personnel. A protocol for reverse transcription loop mediated isothermal amplification (RT-LAMP) has been developed and optimized for the rapid detection of viruses using crude extracts from wheat leaves. The protocol was specific for WSSMV detection, while no reaction was observed with SBCMV or SBWMV, the non-target viruses transmitted by the same vector. The RT-LAMP assay was shown to be as sensitive as the one-step WSSMV specific RT-PCR. The RT-LAMP assay can be performed under field conditions using a portable instrument, and can help the actual spread of WSSMV, an aspect of this virus not yet well understood, to be explored.

MeSH terms

  • Molecular Diagnostic Techniques*
  • Mosaic Viruses*
  • Nucleic Acid Amplification Techniques*
  • Plant Extracts
  • Potyviridae*
  • Triticum

Substances

  • Plant Extracts

Supplementary concepts

  • LAMP assay

Grants and funding

This research was funded by Regione Lombardia, dds n. 4403 28/03/2018, grant n_42— “SURF” and supported through the Euphresco network for phytosanitary research coordination and funding, 2021-A-374: “Diagnosis and epidemiology of viruses infecting cereal crops”. This publication was also supported by the European Virus Archive GLOBAL (EVA-GLOBAL) project that has received funding from the European Union’s Horizon 2020 research and innovation program under grant agreement No. 871029. M.M. was supported by the “SURF” Project and “EVA-GLOBAL” Project. Part of this work was granted by the European Commission – NextGenerationEU, Project SUS-MIRRI.IT "Strengthening the MIRRI Italian Research Infrastructure for Sustainable Bioscience and Bioeconomy”, code n. IR0000005.