Use of electrophoretic techniques and MALDI–TOF MS for rapid and reliable characterization of bacteria: analysis of intact cells, cell lysates, and “washed pellets”

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Publikace nespadá pod Ekonomicko-správní fakultu, ale pod Přírodovědeckou fakultu. Oficiální stránka publikace je na webu muni.cz.
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ŠALPLACHTA Jiří KUBESOVÁ Anna MORAVCOVÁ Dana VYKYDALOVÁ Marie SÜLE Sándor MATOUŠKOVÁ Hana HORKÝ Jaroslav HORKÁ Marie

Rok publikování 2013
Druh Článek v odborném periodiku
Časopis / Zdroj Analytical and Bioanalytical Chemistry
Fakulta / Pracoviště MU

Přírodovědecká fakulta

Citace
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Doi http://dx.doi.org/10.1007/s00216-013-6754-z
Obor Biochemie
Klíčová slova DESORPTION IONIZATION-TIME; FLIGHT MASS-SPECTROMETRY; CAPILLARY-ELECTROPHORESIS; STAPHYLOCOCCUS-AUREUS; HELICOBACTER-PYLORI; CEREBROSPINAL-FLUID; ISOELECTRIC POINT; IDENTIFICATION; MICROORGANISMS; SEPARATION
Popis 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.
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