title: Molecular inversion probe: a new tool for highly specific detection of plant pathogens.
abstract: Highly specific detection methods, capable of reliably identifying plant pathogens are crucial in plant disease management strategies to reduce losses in agriculture by preventing the spread of diseases. We describe a novel molecular inversion probe (MIP) assay that can be potentially developed into a robust multiplex platform to detect and identify plant pathogens. A MIP has been designed for the plant pathogenic fungus Fusarium oxysporum f.sp. conglutinans and the proof of concept for the efficiency of this technology is provided. We demonstrate that this methodology can detect as little as 2.5 ng of pathogen DNA and is highly specific, being able to accurately differentiate Fusarium oxysporum f.sp. conglutinans from other fungal pathogens such as Botrytis cinerea and even pathogens of the same species such as Fusarium oxysporum f.sp. lycopersici. The MIP assay was able to detect the presence of the pathogen in infected Arabidopsis thaliana plants as soon as the tissues contained minimal amounts of pathogen. MIP methods are intrinsically highly multiplexable and future development of specific MIPs could lead to the establishment of a diagnostic method that could potentially screen infected plants for hundreds of pathogens in a single assay.
authors.affiliation: Plant Genetic Engineering Laboratory, School of Agriculture and Food Sciences, University of Queensland, Brisbane, Queensland, Australia; Australian Institute for Bioengineering and Nanotechnology, University of Queensland, Brisbane, Queensland, Australia.
authors.forename: Han Yih
authors.initials: HY
authors.lastname: Lau
authors.affiliation: Australian Institute for Bioengineering and Nanotechnology, University of Queensland, Brisbane, Queensland, Australia.
authors.forename: Ramkumar
authors.initials: R
authors.lastname: Palanisamy
authors.affiliation: Australian Institute for Bioengineering and Nanotechnology, University of Queensland, Brisbane, Queensland, Australia.
authors.forename: Matt
authors.initials: M
authors.lastname: Trau
authors.affiliation: Plant Genetic Engineering Laboratory, School of Agriculture and Food Sciences, University of Queensland, Brisbane, Queensland, Australia.
authors.forename: Jose R
authors.initials: JR
authors.lastname: Botella
mesh_terms: D017360:Arabidopsis / Q000382:microbiology*; D001681:Biological Assay; D004271:DNA, Fungal / Q000235:genetics; D018744:DNA, Plant / Q000378:metabolism; D005670:Fusarium / Q000302:isolation & purification*; D057230:Limit of Detection; D015336:Molecular Probe Techniques; D015335:Molecular Probes / Q000378:metabolism*; D010935:Plant Diseases / Q000382:microbiology*; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D004271:DNA, Fungal; D018744:DNA, Plant; D015335:Molecular Probes