Detection and absolute quantitation of Tomato torrado virus (ToTV) by real time RT-PCR

Tomato torrado virus (ToTV) causes serious damage to the tomato industry and significant economic losses. A quantitative real-time reverse-transcription polymerase chain reaction (RT-qPCR) method using primers and a specific TaqMan (R) MGB probe for ToTV was developed for sensitive detection and qua...

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Main Authors: Herrera-Vásquez, José A., Rubio, Luis, Alfaro-Fernández, Ana, Debreczeni, Diana E., Font-San-Ambrosio, Isabel, Falk, Bryce W., Ferriol, Inmaculada
Format: article
Language:Inglés
Published: 2017
Online Access:http://hdl.handle.net/20.500.11939/4206
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author Herrera-Vásquez, José A.
Rubio, Luis
Alfaro-Fernández, Ana
Debreczeni, Diana E.
Font-San-Ambrosio, Isabel
Falk, Bryce W.
Ferriol, Inmaculada
author_browse Alfaro-Fernández, Ana
Debreczeni, Diana E.
Falk, Bryce W.
Ferriol, Inmaculada
Font-San-Ambrosio, Isabel
Herrera-Vásquez, José A.
Rubio, Luis
author_facet Herrera-Vásquez, José A.
Rubio, Luis
Alfaro-Fernández, Ana
Debreczeni, Diana E.
Font-San-Ambrosio, Isabel
Falk, Bryce W.
Ferriol, Inmaculada
author_sort Herrera-Vásquez, José A.
collection ReDivia
description Tomato torrado virus (ToTV) causes serious damage to the tomato industry and significant economic losses. A quantitative real-time reverse-transcription polymerase chain reaction (RT-qPCR) method using primers and a specific TaqMan (R) MGB probe for ToTV was developed for sensitive detection and quantitation of different ToTV isolates. A standard curve using RNA transcripts enabled absolute quantitation, with a dynamic range from 10(4) to 10(10) ToTV RNA copies/ng of total RNA. The specificity of the RT-qPCR was tested with twenty-three ToTV isolates from tomato (Solanum lycopersicum L), and black nightshade (Solanum nigrum L) collected in Spain, Australia, Hungary and France, which covered the genetic variation range of this virus. This new RT-qPCR assay enables a reproducible, sensitive and specific detection and quantitation of ToTV, which can be a valuable tool in disease management programs and epidemiological studies.
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spelling ReDivia42062025-04-25T14:42:33Z Detection and absolute quantitation of Tomato torrado virus (ToTV) by real time RT-PCR Herrera-Vásquez, José A. Rubio, Luis Alfaro-Fernández, Ana Debreczeni, Diana E. Font-San-Ambrosio, Isabel Falk, Bryce W. Ferriol, Inmaculada Tomato torrado virus (ToTV) causes serious damage to the tomato industry and significant economic losses. A quantitative real-time reverse-transcription polymerase chain reaction (RT-qPCR) method using primers and a specific TaqMan (R) MGB probe for ToTV was developed for sensitive detection and quantitation of different ToTV isolates. A standard curve using RNA transcripts enabled absolute quantitation, with a dynamic range from 10(4) to 10(10) ToTV RNA copies/ng of total RNA. The specificity of the RT-qPCR was tested with twenty-three ToTV isolates from tomato (Solanum lycopersicum L), and black nightshade (Solanum nigrum L) collected in Spain, Australia, Hungary and France, which covered the genetic variation range of this virus. This new RT-qPCR assay enables a reproducible, sensitive and specific detection and quantitation of ToTV, which can be a valuable tool in disease management programs and epidemiological studies. 2017-06-01T10:09:35Z 2017-06-01T10:09:35Z 2015 SEP 1 2015 article Angel Herrera-Vasquez, J., Rubio, L., Alfaro-Fernandez, Ana, Elvira Debreczeni, Diana, Font-San-Ambrosio, I., Falk, Bryce W., Ferriol, I. (2015). Detection and absolute quantitation of Tomato torrado virus (ToTV), by real time RT-PCR. Journal of virological methods, 221, 90-94. 0166-0934 http://hdl.handle.net/20.500.11939/4206 10.1016/j.jviromet.2015.04.029 en openAccess Impreso
spellingShingle Herrera-Vásquez, José A.
Rubio, Luis
Alfaro-Fernández, Ana
Debreczeni, Diana E.
Font-San-Ambrosio, Isabel
Falk, Bryce W.
Ferriol, Inmaculada
Detection and absolute quantitation of Tomato torrado virus (ToTV) by real time RT-PCR
title Detection and absolute quantitation of Tomato torrado virus (ToTV) by real time RT-PCR
title_full Detection and absolute quantitation of Tomato torrado virus (ToTV) by real time RT-PCR
title_fullStr Detection and absolute quantitation of Tomato torrado virus (ToTV) by real time RT-PCR
title_full_unstemmed Detection and absolute quantitation of Tomato torrado virus (ToTV) by real time RT-PCR
title_short Detection and absolute quantitation of Tomato torrado virus (ToTV) by real time RT-PCR
title_sort detection and absolute quantitation of tomato torrado virus totv by real time rt pcr
url http://hdl.handle.net/20.500.11939/4206
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