Real-time multiplex RT-PCR for the simultaneous detection of the five main grapevine viruses

A real-time multiplex RT-PCR has been developed for the simultaneous detection and identification of the major RNA viruses that infect grapevines (Grapevine fanleaf virus, Arabis mosaic virus, Grapevine leafroll-associated virus 1, Grapevine leafroll-associated virus 3 and Grapevine fleck virus). Se...

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Autores principales: López-Fabuel, Irene, Wetzel, Thierry, Bertolini, Edson, Bassler, Alexandra, Vidal, Eduardo, Torres, Luis B., Yuste, Alberto, Olmos, Antonio
Formato: article
Lenguaje:Inglés
Publicado: 2017
Acceso en línea:http://hdl.handle.net/20.500.11939/5528
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author López-Fabuel, Irene
Wetzel, Thierry
Bertolini, Edson
Bassler, Alexandra
Vidal, Eduardo
Torres, Luis B.
Yuste, Alberto
Olmos, Antonio
author_browse Bassler, Alexandra
Bertolini, Edson
López-Fabuel, Irene
Olmos, Antonio
Torres, Luis B.
Vidal, Eduardo
Wetzel, Thierry
Yuste, Alberto
author_facet López-Fabuel, Irene
Wetzel, Thierry
Bertolini, Edson
Bassler, Alexandra
Vidal, Eduardo
Torres, Luis B.
Yuste, Alberto
Olmos, Antonio
author_sort López-Fabuel, Irene
collection ReDivia
description A real-time multiplex RT-PCR has been developed for the simultaneous detection and identification of the major RNA viruses that infect grapevines (Grapevine fanleaf virus, Arabis mosaic virus, Grapevine leafroll-associated virus 1, Grapevine leafroll-associated virus 3 and Grapevine fleck virus). Serial dilutions of infected plant extracts were tested using the new method, and the results were compared with those obtained using a commercially available ELISA and real-time singleplex RT-PCR. The two real-time RT-PCR versions detected up to the same level of dilution and were at least 10,000 times more sensitive than the ELISA. In addition, 158 grapevine plants collected in a survey of the Protected Designation of Origin in Alicante, Spain were compared using the three methods. The results of the molecular methods were very similar, with only four discordant results, and both were able to detect many more infected plants than the. ELISA. The high prevalence of Grapevine fleck virus, Grapevine leafroll-associated virus 3 and Grapevine fanleaf virus suggests that the main pathways of viral introduction are infected plant material that has escaped controls and/or uncontrolled traffic of propagating plant material. Real-time multiplex RT-PCR could be used to facilitate a better control of grapevine viruses. (C) 2012 Elsevier B.V. All rights reserved.
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spelling ReDivia55282025-04-25T14:43:12Z Real-time multiplex RT-PCR for the simultaneous detection of the five main grapevine viruses López-Fabuel, Irene Wetzel, Thierry Bertolini, Edson Bassler, Alexandra Vidal, Eduardo Torres, Luis B. Yuste, Alberto Olmos, Antonio A real-time multiplex RT-PCR has been developed for the simultaneous detection and identification of the major RNA viruses that infect grapevines (Grapevine fanleaf virus, Arabis mosaic virus, Grapevine leafroll-associated virus 1, Grapevine leafroll-associated virus 3 and Grapevine fleck virus). Serial dilutions of infected plant extracts were tested using the new method, and the results were compared with those obtained using a commercially available ELISA and real-time singleplex RT-PCR. The two real-time RT-PCR versions detected up to the same level of dilution and were at least 10,000 times more sensitive than the ELISA. In addition, 158 grapevine plants collected in a survey of the Protected Designation of Origin in Alicante, Spain were compared using the three methods. The results of the molecular methods were very similar, with only four discordant results, and both were able to detect many more infected plants than the. ELISA. The high prevalence of Grapevine fleck virus, Grapevine leafroll-associated virus 3 and Grapevine fanleaf virus suggests that the main pathways of viral introduction are infected plant material that has escaped controls and/or uncontrolled traffic of propagating plant material. Real-time multiplex RT-PCR could be used to facilitate a better control of grapevine viruses. (C) 2012 Elsevier B.V. All rights reserved. 2017-06-01T10:12:31Z 2017-06-01T10:12:31Z 2013 MAR 2013 article Lopez-Fabuel, Irene, Wetzel, Thierry, Bertolini, E., Bassler, Alexandra, Vidal, E., Torres, L. B., Yuste, A., Olmos, A. (2013). Real-time multiplex RT-PCR for the simultaneous detection of the five main grapevine viruses. Journal of virological methods, 188(1-2), 21-24. 0166-0934 http://hdl.handle.net/20.500.11939/5528 10.1016/j.jviromet.2012.11.034 en openAccess Impreso
spellingShingle López-Fabuel, Irene
Wetzel, Thierry
Bertolini, Edson
Bassler, Alexandra
Vidal, Eduardo
Torres, Luis B.
Yuste, Alberto
Olmos, Antonio
Real-time multiplex RT-PCR for the simultaneous detection of the five main grapevine viruses
title Real-time multiplex RT-PCR for the simultaneous detection of the five main grapevine viruses
title_full Real-time multiplex RT-PCR for the simultaneous detection of the five main grapevine viruses
title_fullStr Real-time multiplex RT-PCR for the simultaneous detection of the five main grapevine viruses
title_full_unstemmed Real-time multiplex RT-PCR for the simultaneous detection of the five main grapevine viruses
title_short Real-time multiplex RT-PCR for the simultaneous detection of the five main grapevine viruses
title_sort real time multiplex rt pcr for the simultaneous detection of the five main grapevine viruses
url http://hdl.handle.net/20.500.11939/5528
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