title: Combination of capillary isoelectric focusing in a tapered capillary with MALDI-TOF MS for rapid and reliable identification of Dickeya species from plant samples.
abstract: This study was undertaken to investigate feasibility of a combination of capillary isoelectric focusing (CIEF) in a tapered fused silica (FS) capillary with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) for a rapid and reliable identification of bacteria taken from plant-tissue-containing samples. Eight strains representing different species of the genus Dickeya were selected on the basis of close proximity of their isoelectric points: D. chrysanthemi, D. chrysanthemi bv. parthenii, D. chrysanthemi bv. chrysanthemi, D. dadantii, D. paradisiaca, D. solani, D. diffenbachiae, and D. dianthicola. Because the Dickeya species (spp.) cannot be easily discriminated from each other when CIEF is performed in a cylindrical FS capillary (commonly used in CIEF) even if a narrow pH gradient is used, a tapered FS capillary was employed instead, which enabled satisfactory discrimination of the examined bacteria due to enhanced separation efficiency of CIEF in the tapered FS capillary. CIEF in the tapered FS capillary was also successfully used for the detection and characterization of Dickeya spp. in a plant-tissue-containing sample. Then an off-line combination of CIEF with MALDI-TOF MS was employed for rapid and reliable identification of Dickeya spp. in the plant-tissue-containing sample. It was found that the presence of plant tissue did not affect the results, making the proposed procedure very promising with respect to the fast and reliable detection and identification of bacteria in plant-tissue-containing samples.
authors.affiliation: Institute of Analytical Chemistry of the ASCR, vvi, Brno, Czech Republic. horka@iach.cz
authors.forename: Marie
authors.initials: M
authors.lastname: Horká
authors.forename: Jiří
authors.initials: J
authors.lastname: Salplachta
authors.forename: Pavel
authors.initials: P
authors.lastname: Karásek
authors.forename: Anna
authors.initials: A
authors.lastname: Kubesová
authors.forename: Jaroslav
authors.initials: J
authors.lastname: Horký
authors.forename: Hana
authors.initials: H
authors.lastname: Matoušková
authors.forename: Karel
authors.initials: K
authors.lastname: Slais
authors.forename: Michal
authors.initials: M
authors.lastname: Roth
mesh_terms: D019075:Electrophoresis, Capillary / Q000379:methods; D004755:Enterobacteriaceae / Q000737:chemistry / Q000302:isolation & purification*; D007525:Isoelectric Focusing / Q000379:methods; D010936:Plant Extracts / Q000032:analysis / Q000302:isolation & purification; D010940:Plant Proteins / Q000032:analysis / Q000302:isolation & purification*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods* / Q000592:standards; D013997:Time Factors
chemical_list: D010936:Plant Extracts; D010940:Plant Proteins