Use of electrophoretic techniques and MALDI-TOF MS for rapid and reliable characterization of bacteria: Analysis of intact cells, cell lysates, and "washed pellets"

Jiří Šalplachta, Anna Kubesová, Dana Moravcová, Marie Vykydalová, S. Süle, Hana Matoušková, Jaroslav Horký, Marie Horká

Research output: Contribution to journalArticle

7 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)3165-3175
Number of pages11
JournalAnalytical and Bioanalytical Chemistry
Volume405
Issue number10
DOIs
Publication statusPublished - Apr 2013

Fingerprint

Ionization
Mass spectrometry
Desorption
Mass Spectrometry
Bacteria
Lasers
Isoelectric Focusing
Rhizobium leguminosarum
Capillary Electrophoresis
Gels
Capillary electrophoresis
Rhizobiaceae
Electrophoresis
Sinorhizobium meliloti
Microorganisms
Dermatoglyphics
DNA

Keywords

  • Bacteria
  • Characterization of bacteria
  • CIEF
  • CZE
  • IEF
  • MALDI

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry

Cite this

Use of electrophoretic techniques and MALDI-TOF MS for rapid and reliable characterization of bacteria : Analysis of intact cells, cell lysates, and "washed pellets". / Šalplachta, Jiří; Kubesová, Anna; Moravcová, Dana; Vykydalová, Marie; Süle, S.; Matoušková, Hana; Horký, Jaroslav; Horká, Marie.

In: Analytical and Bioanalytical Chemistry, Vol. 405, No. 10, 04.2013, p. 3165-3175.

Research output: Contribution to journalArticle

Šalplachta, Jiří ; Kubesová, Anna ; Moravcová, Dana ; Vykydalová, Marie ; Süle, S. ; Matoušková, Hana ; Horký, Jaroslav ; Horká, Marie. / Use of electrophoretic techniques and MALDI-TOF MS for rapid and reliable characterization of bacteria : Analysis of intact cells, cell lysates, and "washed pellets". In: Analytical and Bioanalytical Chemistry. 2013 ; Vol. 405, No. 10. pp. 3165-3175.
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