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

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

van Brunschot, S.L. ORCID: 0000-0002-9634-9463, Bergervoet, J.H.W., Pagendam, D.E., de Weerdt, M., Geering, A.D.W., Drenth, A. and van der Vlugt, R.A.A. (2014) A bead-based suspension array for the multiplexed detection of begomoviruses and their whitefly vectors. Journal of Virological Methods, 198. pp. 86-94. ISSN 0166-0934 (Print), 1879-0984 (Online) (doi:10.1016/j.jviromet.2013.12.014)

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Abstract

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 200 fg 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.

Item Type: Article
Uncontrolled Keywords: Luminex; Diagnostic; Bemisia tabaci; Tomato yellow leaf curl virus; qPCR
Subjects: S Agriculture > S Agriculture (General)
Faculty / Department / 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 15:07
Selected for GREAT 2016: None
Selected for GREAT 2017: GREAT c
Selected for GREAT 2018: None
URI: http://gala.gre.ac.uk/id/eprint/17208

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