Detection and absolute quantitation of Broad bean wilt virus 1 (BBWV-1) and BBWV-2 by real time RT-PCR

Broad bean wilt virus 1 (BBWV-1) and BBWV-2 are the two most significant viruses in the genus Fabavirus, causing damage to many economically important agricultural crops worldwide. A quantitative real-time reverse transcription-polymerase chain reaction (RT-qPCR) procedure using two TaqMan (R) MGB p...

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Main Authors: Ferriol, Inmaculada, Ruiz-Ruiz, Susana, Rubio, Luis
Format: Artículo
Language:Inglés
Published: 2017
Online Access:http://hdl.handle.net/20.500.11939/5203
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author Ferriol, Inmaculada
Ruiz-Ruiz, Susana
Rubio, Luis
author_browse Ferriol, Inmaculada
Rubio, Luis
Ruiz-Ruiz, Susana
author_facet Ferriol, Inmaculada
Ruiz-Ruiz, Susana
Rubio, Luis
author_sort Ferriol, Inmaculada
collection ReDivia
description Broad bean wilt virus 1 (BBWV-1) and BBWV-2 are the two most significant viruses in the genus Fabavirus, causing damage to many economically important agricultural crops worldwide. A quantitative real-time reverse transcription-polymerase chain reaction (RT-qPCR) procedure using two TaqMan (R) MGB probes was developed for sensitive and specific detection and quantitation of BBWV-1 and BBWV-2. Primers and probes were designed from conserved sequence stretches to detect all isolates of each virus. Standard curves using RNA transcripts identical to both TaqMan (R) MGB probes enabled absolute quantitation, with a wide dynamic range and high sensitivity (10(3)-10(10) RNA molecules). RT-qPCR was assayed with genetically divergent BBWV-1 and BBWV-2 isolates from different plant hosts and countries, and was used to evaluate the temporal accumulation of BBWV-1 RNA in two plant hosts. (C) 2011 Elsevier B.V. All rights reserved.
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spelling ReDivia52032025-04-25T14:45:38Z Detection and absolute quantitation of Broad bean wilt virus 1 (BBWV-1) and BBWV-2 by real time RT-PCR Ferriol, Inmaculada Ruiz-Ruiz, Susana Rubio, Luis Broad bean wilt virus 1 (BBWV-1) and BBWV-2 are the two most significant viruses in the genus Fabavirus, causing damage to many economically important agricultural crops worldwide. A quantitative real-time reverse transcription-polymerase chain reaction (RT-qPCR) procedure using two TaqMan (R) MGB probes was developed for sensitive and specific detection and quantitation of BBWV-1 and BBWV-2. Primers and probes were designed from conserved sequence stretches to detect all isolates of each virus. Standard curves using RNA transcripts identical to both TaqMan (R) MGB probes enabled absolute quantitation, with a wide dynamic range and high sensitivity (10(3)-10(10) RNA molecules). RT-qPCR was assayed with genetically divergent BBWV-1 and BBWV-2 isolates from different plant hosts and countries, and was used to evaluate the temporal accumulation of BBWV-1 RNA in two plant hosts. (C) 2011 Elsevier B.V. All rights reserved. 2017-06-01T10:11:53Z 2017-06-01T10:11:53Z 2011 NOV 2011 article Ferriol, I., Ruiz-Ruiz, S., Rubio, L. (2011). Detection and absolute quantitation of Broad bean wilt virus 1 (BBWV-1), and BBWV-2 by real time RT-PCR. Journal of virological methods, 177(2), 202-205. 0166-0934 http://hdl.handle.net/20.500.11939/5203 http://hdl.handle.net/20.500.11939/5203 10.1016/j.jviromet.2011.08.003 en openAccess Impreso
spellingShingle Ferriol, Inmaculada
Ruiz-Ruiz, Susana
Rubio, Luis
Detection and absolute quantitation of Broad bean wilt virus 1 (BBWV-1) and BBWV-2 by real time RT-PCR
title Detection and absolute quantitation of Broad bean wilt virus 1 (BBWV-1) and BBWV-2 by real time RT-PCR
title_full Detection and absolute quantitation of Broad bean wilt virus 1 (BBWV-1) and BBWV-2 by real time RT-PCR
title_fullStr Detection and absolute quantitation of Broad bean wilt virus 1 (BBWV-1) and BBWV-2 by real time RT-PCR
title_full_unstemmed Detection and absolute quantitation of Broad bean wilt virus 1 (BBWV-1) and BBWV-2 by real time RT-PCR
title_short Detection and absolute quantitation of Broad bean wilt virus 1 (BBWV-1) and BBWV-2 by real time RT-PCR
title_sort detection and absolute quantitation of broad bean wilt virus 1 bbwv 1 and bbwv 2 by real time rt pcr
url http://hdl.handle.net/20.500.11939/5203
http://hdl.handle.net/20.500.11939/5203
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