title: Field Demonstration of a Multiplexed Point-of-Care Diagnostic Platform for Plant Pathogens.
abstract: Effective disease management strategies to prevent catastrophic crop losses require rapid, sensitive, and multiplexed detection methods for timely decision making. To address this need, a rapid, highly specific and sensitive point-of-care method for multiplex detection of plant pathogens was developed by taking advantage of surface-enhanced Raman scattering (SERS) labeled nanotags and recombinase polymerase amplification (RPA), which is a rapid isothermal amplification method with high specificity. In this study, three agriculturally important plant pathogens (Botrytis cinerea, Pseudomonas syringae, and Fusarium oxysporum) were used to demonstrate potential translation into the field. The RPA-SERS method was faster, more sensitive than polymerase chain reaction, and could detect as little as 2 copies of B. cinerea DNA. Furthermore, multiplex detection of the three pathogens was demonstrated for complex systems such as the Arabidopsis thaliana plant and commercial tomato crops. To demonstrate the potential for on-site field applications, a rapid single-tube RPA/SERS assay was further developed and successfully performed for a specific target outside of a laboratory setting.
authors.affiliation: Centre for Personalized Nanomedicine, Australian Institute for Bioengineering and Nanotechnology, The University of Queensland , Brisbane QLD 4072, Australia.|Plant Genetic Engineering Laboratory, School of Agriculture and Food Sciences, The University of Queensland , Brisbane QLD 4072, Australia.
authors.forename: Han Yih
authors.initials: HY
authors.lastname: Lau
authors.affiliation: Centre for Personalized Nanomedicine, Australian Institute for Bioengineering and Nanotechnology, The University of Queensland , Brisbane QLD 4072, Australia.
authors.forename: Yuling
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Centre for Personalized Nanomedicine, Australian Institute for Bioengineering and Nanotechnology, The University of Queensland , Brisbane QLD 4072, Australia.
authors.forename: Eugene J H
authors.initials: EJ
authors.lastname: Wee
authors.affiliation: Plant Genetic Engineering Laboratory, School of Agriculture and Food Sciences, The University of Queensland , Brisbane QLD 4072, Australia.
authors.forename: Jose R
authors.initials: JR
authors.lastname: Botella
authors.affiliation: Centre for Personalized Nanomedicine, Australian Institute for Bioengineering and Nanotechnology, The University of Queensland , Brisbane QLD 4072, Australia.|School of Chemistry and Molecular Biosciences, The University of Queensland , Brisbane QLD 4072, Australia.
authors.forename: Matt
authors.initials: M
authors.lastname: Trau
mesh_terms: D017360:Arabidopsis / Q000382:microbiology; D020171:Botrytis / Q000235:genetics / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D005670:Fusarium / Q000235:genetics / Q000302:isolation & purification*; D006046:Gold; D018551:Solanum lycopersicum / Q000382:microbiology; D053768:Metal Nanoparticles / Q000737:chemistry; D021141:Nucleic Acid Amplification Techniques; D010935:Plant Diseases / Q000382:microbiology*; D018515:Plant Leaves / Q000235:genetics; D019095:Point-of-Care Systems; D044224:Pseudomonas syringae / Q000235:genetics / Q000302:isolation & purification; D013059:Spectrum Analysis, Raman
chemical_list: D004269:DNA, Bacterial; D004271:DNA, Fungal; D006046:Gold