Exploiting generic platform technologies for the detection and identification of plant pathogens
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The detection and identification of plant pathogens currently relies upon a very diverse range of techniques and skills, from traditional culturing and taxonomic skills to modern molecular-based methods. The wide range of methods employed reflects the great diversity of plant pathogens and the hosts they infect. The well-documented decline in taxonomic expertise, along with the need to develop ever more rapid and sensitive diagnostic methods has provided an impetus to develop technologies that are both generic and able to complement traditional skills and techniques. Real-time polymerase chain reaction (PCR) is emerging as one such generic platform technology and one that is well suited to high-throughput detection of a limited number of known target pathogens. Real-time PCR is now exploited as a front line diagnostic screening tool in human health, animal health, homeland security, biosecurity as well as plant health. Progress with developing generic techniques for plant pathogen identification, particularly of unknown samples, has been less rapid. Diagnostic microarrays and direct nucleic acid sequencing (de novo sequencing) both have potential as generic methods for the identification of unknown plant pathogens but are unlikely to be suitable as high-throughput detection techniques. This paper will review the application of generic technologies in the routine laboratory as well as highlighting some new techniques and the trend towards multi-disciplinary studies.
KeywordsPlant health Diagnostics Detection Real-time PCR DNA barcoding Microarrays Isothermal amplification LAMP Direct tuber testing Molecular diagnostics TaqMan De novo sequencing Pyrosequencing
The authors would like to acknowledge funding from Plant Health Division and Chief Scientist Group of Defra (www.bio-chip.co.uk), and also the European Union for funding under the fifth framework programme (www.diagchip.co.uk) and also the sixth framework programme (www.portcheck.eu.com) project (SSPE-CT-2004-502348). In addition the authors would like to acknowledge the help and support provided through the COST project ‘Agricultural Biomarkers for Array Technology’ (http://www.cost853.ch/).
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