Use of hybridization melting kinetics for detecting Phytophthora species using three-dimensional microarrays: demonstration of a novel concept for the differentiation of detection targets

N. Anderson; M. Szemes; P.A. O'Brien; M. De Weerdt; C. Schoen; P. Boender; P. Bonants

doi:10.1016/j.mycres.2006.04.003

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Use of hybridization melting kinetics for detecting Phytophthora species using three-dimensional microarrays: demonstration of a novel concept for the differentiation of detection targets

Journal article

Peer reviewed

Use of hybridization melting kinetics for detecting Phytophthora species using three-dimensional microarrays: demonstration of a novel concept for the differentiation of detection targets

N. Anderson, M. Szemes, P.A. O'Brien, M. De Weerdt, C. Schoen, P. Boender and P. Bonants

Mycological Research, Vol.110(6), pp.664-671

2006

DOI: https://doi.org/10.1016/j.mycres.2006.04.003

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Abstract

Microarray-based detection is limited by variable and inconsistent hybridization intensities across the diversity of probes used in each array. In this paper, we introduce a novel concept for the differentiation of detection targets using duplex melting kinetics. A microarray assay was developed on a PamChip microarray enabling the differentiation of target Phytophthora species using the melting kinetics of probe-target duplexes. In the majority of cases the hybridization kinetics of target and non-target duplexes differed significantly. Analysis of the melting kinetics of duplexes formed by probes with target and non-target DNA was found to be an effective method for determining specific hybridization and was independent of fluctuations in hybridization signal intensity. This form of analysis was more robust than the traditional approach based on hybridization intensity, and enabled the detection of individual Phytophthora species and mixtures thereof.

Details

Title: Use of hybridization melting kinetics for detecting Phytophthora species using three-dimensional microarrays: demonstration of a novel concept for the differentiation of detection targets
Authors/Creators: N. Anderson (Author/Creator) - Murdoch University
M. Szemes (Author/Creator) - Research International (United States)
P.A. O'Brien (Author/Creator) - Murdoch University
M. De Weerdt (Author/Creator) - Research International (United States)
C. Schoen (Author/Creator) - Research International (United States)
P. Boender (Author/Creator) - PamGene (Netherlands)
P. Bonants (Author/Creator)
Publication Details: Mycological Research, Vol.110(6), pp.664-671
Publisher: Elsevier
Identifiers: 991005540643107891
Copyright: (c) Elsevier
Murdoch Affiliation: Centre for Phytophthora Science and Management
Language: English
Resource Type: Journal article

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Collaboration types: Industry collaboration; Domestic collaboration; International collaboration
Citation topics: 3 Agriculture, Environment & Ecology; 3.97 Plant Pathology; 3.97.1439 Phytophthora Management
Web Of Science research areas: Mycology
ESI research areas: Plant & Animal Science