title: Use of electrophoretic techniques and MALDI-TOF MS for rapid and reliable characterization of bacteria: analysis of intact cells, cell lysates, and "washed pellets".
abstract: In this study electrophoretic and mass spectrometric analysis of three types of bacterial sample (intact cells, cell lysates, and "washed pellets") were used to develop an effective procedure for the characterization of bacteria. The samples were prepared from specific bacterial strains. Five strains representing different species of the family Rhizobiaceae were selected as model microorganisms: Rhizobium leguminosarum bv. trifolii, R. leguminosarum bv. viciae, R. galegae, R. loti, and Sinorhizobium meliloti. Samples of bacteria were subjected to analysis by four techniques: capillary zone electrophoresis (CZE), capillary isoelectric focusing (CIEF), gel IEF, and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). These methods are potential alternatives to DNA-based methods for rapid and reliable characterization of bacteria. Capillary electrophoretic (CZE and CIEF) analysis of intact cells was suitable for characterization of different bacterial species. CIEF fingerprints of "washed pellets" and gel IEF of cell lysates helped to distinguish between closely related bacterial species that were not sufficiently differentiated by capillary electrophoretic analysis of intact cells. MALDI-TOF MS of "washed pellets" enabled more reliable characterization of bacteria than analysis of intact cells or cell lysates. Electrophoretic techniques and MALDI-TOF MS can both be successfully used to complement standard methods for rapid characterization of bacteria.
authors: Institute of Analytical Chemistry of the ASCR, v v i, Brno, Czech Republic. salplachta@iach.cz
authors: Jiří
authors: J
authors: Salplachta
authors: Anna
authors: A
authors: Kubesová
authors: Dana
authors: D
authors: Moravcová
authors: Marie
authors: M
authors: Vykydalová
authors: Sándor
authors: S
authors: Süle
authors: Hana
authors: H
authors: Matoušková
authors: Jaroslav
authors: J
authors: Horký
authors: Marie
authors: M
authors: Horká
mesh_terms: D019075:Electrophoresis, Capillary / Q000379:methods*; D007525:Isoelectric Focusing / Q000379:methods*; D050296:Microbial Viability; D012231:Rhizobium / Q000737:chemistry* / Q000145:classification / Q000302:isolation & purification; D016962:Sinorhizobium meliloti / Q000737:chemistry* / Q000145:classification / Q000302:isolation & purification; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*