van Brunschot, Sharon L. ORCID: 0000-0002-9634-9463, Bergervoet, Jan H. W., Pagendam, Daniel E., de Weerdt, Marjanne, Geering, Andrew D. W., Drenth, André and van der Vlugt, René A. A. (2014) Development of a multiplexed bead-based suspension array for the detection and discrimination of Pospiviroid plant pathogens. PLoS ONE, 9 (1):e84743. ISSN 1932-6203 (doi:https://doi.org/10.1371/journal.pone.0084743)

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Abstract

Efficient and reliable diagnostic tools for the routine indexing and certification of clean propagating material are essential for the management of pospiviroid diseases in horticultural crops. This study describes the development of a true multiplexed diagnostic method for the detection and identification of all nine currently recognized pospiviroid species in one assay using Luminex bead-based suspension array technology. In addition, a new data-driven, statistical method is presented for establishing thresholds for positivity for individual assays within multiplexed arrays. When applied to the multiplexed array data generated in this study, the new method was shown to have better control of false positives and false negative results than two other commonly used approaches for setting thresholds. The 11-plex Luminex MagPlex-TAG pospiviroid array described here has a unique hierarchical assay design, incorporating a near-universal assay in addition to nine species-specific assays, and a co-amplified plant internal control assay for quality assurance purposes. All assays of the multiplexed array were shown to be 100% specific, sensitive and reproducible. The multiplexed array described herein is robust, easy to use, displays unambiguous results and has strong potential for use in routine pospiviroid indexing to improve disease management strategies.

Item Type:	Article
Additional Information:	Copyright: © 2014 van Brunschot et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Uncontrolled Keywords:	Plant Pathogen; Viroid; Diagnostic Test; Multiplex; Quarantine; Luminex; Bead array
Subjects:	S Agriculture > S Agriculture (General)
Faculty / School / Research Centre / Research Group:	Faculty of Engineering & Science Faculty of Engineering & Science > Natural Resources Institute Faculty of Engineering & Science > Natural Resources Institute > Agriculture, Health & Environment Department Faculty of Engineering & Science > Natural Resources Institute > Plant Health Research Group
Last Modified:	01 Jun 2017 12:05
URI:	http://gala.gre.ac.uk/id/eprint/17172

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