Development of simplex and multiplex RT-qPCR assays for the detection of three cryptic viruses of black-grass (Alopecurus myosuroides).


Journal

Journal of virological methods
ISSN: 1879-0984
Titre abrégé: J Virol Methods
Pays: Netherlands
ID NLM: 8005839

Informations de publication

Date de publication:
02 2022
Historique:
received: 09 03 2021
revised: 17 06 2021
accepted: 26 11 2021
pubmed: 2 12 2021
medline: 22 3 2022
entrez: 1 12 2021
Statut: ppublish

Résumé

Simplex and multiplex RT-qPCR assays were developed for Alopecurus myosuroides partitivirus 1 (AMPV1), Alopecurus myosuroides partitivirus 2 (AMPV2) and Alopecurus myosuroides varicosavirus 1 (AMVV1), and compared to the existing conventional PCR assays. All assays had a high specificity and their sensitivity was increased compared to the conventional RT-PCR assays. As viral quantification is an important element in comparative experiments, the effect of high- and low-temperature drying treatments, prior to RNA extraction and analysis, was studied and optimised. AMVV1 detection was reduced by both drying treatments, but particularly by the high-temperature. AMPV1 and AMPV2 detection on the other hand was not impeded by the drying treatments, and enables standardisation of plant tissue prior to extraction, in particular for quantitative analysis.

Identifiants

pubmed: 34848282
pii: S0166-0934(21)00328-1
doi: 10.1016/j.jviromet.2021.114389
pii:
doi:

Substances chimiques

Herbicides 0

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

114389

Informations de copyright

Copyright © 2021 Elsevier B.V. All rights reserved.

Auteurs

Jone Santín Azcona (J)

Fera Science Ltd.York Biotech Campus, Sand Hutton, York, YO41 1LZ United Kingdom. Electronic address: jonesantin@gmail.com.

Adrian Fox (A)

Fera Science Ltd.York Biotech Campus, Sand Hutton, York, YO41 1LZ United Kingdom. Electronic address: Adrian.Fox@fera.co.uk.

Neil Boonham (N)

Newcastle University, Newcastle upon Tyne, NE1 7RU, United Kingdom. Electronic address: Neil.Boonham@newcastle.ac.uk.

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Classifications MeSH