A bead-based suspension array for the multiplexed detection of begomoviruses and their whitefly vectors.

Autor: van Brunschot SL; Plant Biosecurity Cooperative Research Centre, LPO Box 5012, Bruce, ACT 2617, Australia; School of Agriculture and Food Sciences, The University of Queensland, St. Lucia, QLD 4072, Australia., Bergervoet JH; Plant Research International, Wageningen University and Research Centre, PO Box 16, 6700 AA Wageningen, The Netherlands., Pagendam DE; CSIRO Mathematics, Informatics and Statistics, Ecosciences Precinct, Dutton Park, QLD 4102, Australia., de Weerdt M; Plant Research International, Wageningen University and Research Centre, PO Box 16, 6700 AA Wageningen, The Netherlands., Geering AD; Centre for Plant Science, Queensland Alliance for Agriculture and Food Innovation, The University of Queensland, GPO Box 267, Brisbane, QLD 4001, Australia., Drenth A; Centre for Plant Science, Queensland Alliance for Agriculture and Food Innovation, The University of Queensland, GPO Box 267, Brisbane, QLD 4001, Australia., van der Vlugt RA; Plant Research International, Wageningen University and Research Centre, PO Box 16, 6700 AA Wageningen, The Netherlands. Electronic address: rene.vandervlugt@wur.nl.
Jazyk: angličtina
Zdroj: Journal of virological methods [J Virol Methods] 2014 Mar; Vol. 198, pp. 86-94. Date of Electronic Publication: 2013 Dec 31.
DOI: 10.1016/j.jviromet.2013.12.014
Abstrakt: Bead-based suspension array systems enable simultaneous fluorescence-based identification of multiple nucleic acid targets in a single reaction. This study describes the development of a novel approach to plant virus and vector diagnostics, a multiplexed 7-plex array that comprises a hierarchical set of assays for the simultaneous detection of begomoviruses and Bemisia tabaci, from both plant and whitefly samples. The multiplexed array incorporates genus, species and strain-specific assays, offering a unique approach for identifying both known and unknown viruses and B. tabaci species. When tested against a large panel of sequence-characterized begomovirus and whitefly samples, the array was shown to be 100% specific to the homologous target. Additionally, the multiplexed array was highly sensitive, efficiently and concurrently determining both virus and whitefly identity from single viruliferous whitefly samples. The detection limit for one assay within the multiplexed array that specifically detects Tomato yellow leaf curl virus-Israel (TYLCV-IL) was quantified as 200fg of TYLCV-IL DNA, directly equivalent to that of TYLCV-specific qPCR. Highly reproducible results were obtained over multiple tests. The flexible multiplexed array described in this study has great potential for use in plant quarantine, biosecurity and disease management programs worldwide.
(Copyright © 2014 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
Externí odkaz: