title: Label-free detection of Phytophthora ramorum using surface-enhanced Raman spectroscopy.
abstract: In this study, we report on a novel approach for the label-free and species-specific detection of the plant pathogen Phytophthora ramorum from real samples using surface enhanced Raman scattering (SERS). In this context, we consider the entire analysis chain including sample preparation, DNA isolation, amplification and hybridization on SERS substrate-immobilized adenine-free capture probes. Thus, the SERS-based detection of target DNA is verified by the strong spectral feature of adenine which indicates the presence of hybridized target DNA. This property was realized by replacing adenine moieties in the species-specific capture probes with 2-aminopurine. In the case of the matching capture and target sequence, the characteristic adenine peak serves as an indicator for specific DNA hybridization. Altogether, this is the first assay demonstrating the detection of a plant pathogen from an infected plant material by label-free SERS employing DNA hybridization on planar SERS substrates consisting of silver nanoparticles.
authors.affiliation: Leibniz Institute of Photonic Technology Jena (IPHT), Albert-Einstein-Straße 9, 07745 Jena, Germany. karina.weber@ipht-jena.de.
authors.forename: Sezin
authors.initials: S
authors.lastname: Yüksel
authors.forename: Lydia
authors.initials: L
authors.lastname: Schwenkbier
authors.forename: Sibyll
authors.initials: S
authors.lastname: Pollok
authors.forename: Karina
authors.initials: K
authors.lastname: Weber
authors.forename: Dana
authors.initials: D
authors.lastname: Cialla-May
authors.forename: Jürgen
authors.initials: J
authors.lastname: Popp
mesh_terms: D015075:2-Aminopurine / Q000737:chemistry; D000225:Adenine / Q000737:chemistry; D004247:DNA / Q000737:chemistry*; D053768:Metal Nanoparticles / Q000737:chemistry; D008855:Microscopy, Electron, Scanning; D036103:Nanotechnology; D009693:Nucleic Acid Hybridization; D010838:Phytophthora / Q000302:isolation & purification*; D018515:Plant Leaves / Q000382:microbiology; D029793:Rhododendron / Q000382:microbiology; D012542:Scattering, Radiation; D012834:Silver / Q000737:chemistry; D013059:Spectrum Analysis, Raman / Q000379:methods*; D013499:Surface Properties
chemical_list: D012834:Silver; D015075:2-Aminopurine; D004247:DNA; D000225:Adenine