Articles
ISOTHERMAL AMPLIFICATION FOR DETECTION OF PLUM POX VIRUS
Article number
781_31
Pages
209 – 214
Language
English
Abstract
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.
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.
Publication
Authors
A. Olmos, E. Bertolini, M. Cambra
Keywords
NASBA, flow-through hybridisation
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