Optimisation of intact cell MALDI method for fingerprinting of methicillin-resistant Staphylococcus aureus

Kathryn A. Jackson*, V. Edwards-Jones, C. W. Sutton, A. J. Fox

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

91 Citations (Scopus)

Abstract

The use of matrix-assisted laser desorption/ionisation time-of-flight (MALDI-TOF) mass spectrometry on intact cell microorganisms, Intact Cell MALDI (ICM), has been shown by numerous workers to yield effective species level identification. Early work highlighted the significant effect that variation in culture media, incubation conditions and length of incubation had on the spectra produced. Therefore, in order to achieve reliable and reproducible species level identification and sub-typing of microorganisms from ICM fingerprints, it has been essential to develop standardised methods. For methicillin-resistant Staphylococcus aureus (MRSA), a major nosocomial pathogen, we have developed such a standardised method. In this paper we present the experimental parameters, namely, the incubation period, the number of passages required from lyophilised or stored isolates, the method of deposition of the bacterial cells, the concentration of matrix solution, the drying time of bacterial cells prior to the addition of the matrix solution, the time between preparation of the bacterial/matrix sample and analysis and the MALDI pulsed extraction setting, which were considered during the development of defined methods.

Original languageEnglish
Pages (from-to)273-284
Number of pages12
JournalJournal of Microbiological Methods
Volume62
Issue number3 SPEC. ISS.
DOIs
Publication statusPublished - Sep 2005

Keywords

  • ICM
  • Intact Cell MALDI
  • MRSA

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