Isothermal amplification for detection of Plum pox virus

A nucleic acid sequence-based amplification method coupled with flow-through hybridisation (NASBA-FH) was developed for Plum pox virus (PPV) detection. The detection limit of the NASBA-FH was established at two copies of control transcripts, resulting 10 times higher than that obtained by Co-PCR and...

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Main Authors: Olmos, Antonio, Bertolini, Edson, Cambra, Mariano
Other Authors: Ertunc, F.
Format: Objeto de conferencia
Language:Inglés
Published: 2017
Online Access:http://hdl.handle.net/20.500.11939/4254
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author Olmos, Antonio
Bertolini, Edson
Cambra, Mariano
author2 Ertunc, F.
author_browse Bertolini, Edson
Cambra, Mariano
Ertunc, F.
Olmos, Antonio
author_facet Ertunc, F.
Olmos, Antonio
Bertolini, Edson
Cambra, Mariano
author_sort Olmos, Antonio
collection ReDivia
description A nucleic acid sequence-based amplification method coupled with flow-through hybridisation (NASBA-FH) was developed for Plum pox virus (PPV) detection. The detection limit of the NASBA-FH was established at two copies of control transcripts, resulting 10 times higher than that obtained by Co-PCR and 1000 times higher than the sensitivity afforded by RT-PCR. In addition, samples from 253 stone-fruit trees were collected during winter and analysed. The samples were tested using methods recommended by the European and Mediterranean Plant Protection Organization to detect PPV (DASI-ELISA, RT-PCR and Co-PCR) and by NASBA-FH. PPV diagnosis by ELISA and NASBA-FH coincided in 90.5%, while ELISA and PCR-based methods coincided in the diagnosis of 91.3% of the trees and PCR-based methods with NASBA-FH agreed in 95.2%. Results support that NASBA-FH is a suitable molecular method for routine PPV detection in the winter period.
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institution Instituto Valenciano de Investigaciones Agrarias (IVIA)
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spelling ReDivia42542025-04-25T14:51:35Z Isothermal amplification for detection of Plum pox virus ACTA HORTICULTURAE Olmos, Antonio Bertolini, Edson Cambra, Mariano Ertunc, F. Caglayan, Kadriye A nucleic acid sequence-based amplification method coupled with flow-through hybridisation (NASBA-FH) was developed for Plum pox virus (PPV) detection. The detection limit of the NASBA-FH was established at two copies of control transcripts, resulting 10 times higher than that obtained by Co-PCR and 1000 times higher than the sensitivity afforded by RT-PCR. In addition, samples from 253 stone-fruit trees were collected during winter and analysed. The samples were tested using methods recommended by the European and Mediterranean Plant Protection Organization to detect PPV (DASI-ELISA, RT-PCR and Co-PCR) and by NASBA-FH. PPV diagnosis by ELISA and NASBA-FH coincided in 90.5%, while ELISA and PCR-based methods coincided in the diagnosis of 91.3% of the trees and PCR-based methods with NASBA-FH agreed in 95.2%. Results support that NASBA-FH is a suitable molecular method for routine PPV detection in the winter period. 2017-06-01T10:09:41Z 2017-06-01T10:09:41Z 2008 2008 conferenceObject Olmos, A., Bertolini, E., Cambra, M. (2008). Isothermal amplification for detection of Plum pox virus. Proceedings of the Twentieth International Symposium on Virus and Virus-Like Diseases of Temperate Fruit Crops - Fruit Tree Diseases, (781), 209-213. 0567-7572; 978-90-6605-080-8 http://hdl.handle.net/20.500.11939/4254 10.17660/ActaHortic.2008.781.31 en openAccess Impreso
spellingShingle Olmos, Antonio
Bertolini, Edson
Cambra, Mariano
Isothermal amplification for detection of Plum pox virus
title Isothermal amplification for detection of Plum pox virus
title_full Isothermal amplification for detection of Plum pox virus
title_fullStr Isothermal amplification for detection of Plum pox virus
title_full_unstemmed Isothermal amplification for detection of Plum pox virus
title_short Isothermal amplification for detection of Plum pox virus
title_sort isothermal amplification for detection of plum pox virus
url http://hdl.handle.net/20.500.11939/4254
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