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Quantitative multiplex detection of plant pathogens using a novel ligation probe-based system coupled with universal, high-throughput real-time PCR on OpenArraysTM

Van Doorn, R. and Szemes, M. and Bonants, P.J.M. and Kowalchuk, G.A. and Sallas, J. and Ortenberg, E. and Schoen, C.D. (2007) Quantitative multiplex detection of plant pathogens using a novel ligation probe-based system coupled with universal, high-throughput real-time PCR on OpenArraysTM. BMC Genomics, 8, 276-. ISSN 1471-2164.

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Official URL: http://dx.doi.org/10.1186/1471-2164-8-276

Abstract

Background Diagnostics and disease-management strategies require technologies to enable the simultaneous detection and quantification of a wide range of pathogenic microorganisms. Most multiplex, quantitative detection methods available suffer from compromises between the level of multiplexing, throughput and accuracy of quantification. Here, we demonstrate the efficacy of a novel, high-throughput, ligation-based assay for simultaneous quantitative detection of multiple plant pathogens. The ligation probes Results The performance of the PRI-lock detection system was demonstrated using 13 probes targeting several significant plant pathogens at different taxonomic levels. All probes specifically detected their corresponding targets and provided perfect discrimination against non-target organisms with very similar ligation target sites. The nucleic acid targets could be reliably quantified over 5 orders of magnitude with a dynamic detection range of more than 104. Pathogen quantification was equally robust i Conclusion This novel assay enables very specific, high-throughput, quantitative detection of multiple pathogens over a wide range of target concentrations and should be easily adaptable for versatile diagnostic purposes.

Item Type:Article
Institutes:Nederlands Instituut voor Ecologie (NIOO)
ID Code:4620
Deposited On:16 Sep 2009 02:00
Last Modified:02 Nov 2011 16:37

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