Field Demonstration of a Multiplexed Point-of-Care Diagnostic Platform for Plant Pathogens

Anal Chem. 2016 Aug 16;88(16):8074-81. doi: 10.1021/acs.analchem.6b01551. Epub 2016 Jul 27.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis / microbiology
  • Botrytis / genetics
  • Botrytis / isolation & purification*
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / genetics
  • DNA, Bacterial / metabolism
  • DNA, Fungal / chemistry
  • DNA, Fungal / genetics
  • DNA, Fungal / metabolism
  • Fusarium / genetics
  • Fusarium / isolation & purification*
  • Gold
  • Metal Nanoparticles / chemistry
  • Nucleic Acid Amplification Techniques
  • Plant Diseases / microbiology*
  • Plant Leaves / genetics
  • Point-of-Care Systems*
  • Pseudomonas syringae / genetics
  • Pseudomonas syringae / isolation & purification
  • Solanum lycopersicum / microbiology
  • Spectrum Analysis, Raman

Substances

  • DNA, Bacterial
  • DNA, Fungal
  • Gold