Optimizing identification of clinically relevant gram-positive organisms by use of the bruker biotyper matrix-assisted laser desorption ionization-time of flight mass spectrometry system

Erin McElvania TeKippe, Sunni Shuey, David W. Winkler, Meghan A. Butler, Carey Ann D Burnham

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Abstract

Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) can be used as a method for the rapid identification of microorganisms. This study evaluated the Bruker Biotyper (MALDI-TOF MS) system for the identification of clinically relevant Gram-positive organisms. We tested 239 aerobic Gram-positive organisms isolated from clinical specimens. We evaluated 4 direct-smear methods, including "heavy" (H) and "light" (L) smears, with and without a 1-=l direct formic acid (FA) overlay. The quality measure assigned to a MALDI-TOF MS identification is a numerical value or "score." We found that a heavy smear with a formic acid overlay (H+FA) produced optimal MALDI-TOF MS identification scores and the highest percentage of correctly identified organisms. Using a score of>2.0, we identified 183 of the 239 isolates (76.6%) to the genus level, and of the 181 isolates resolved to the species level, 141 isolates (77.9%) were correctly identified. To maximize the number of correct identifications while minimizing misidentifications, the data were analyzed using a score of>1.7 for genus- and species-level identification. Using this score, 220 of the 239 isolates (92.1%) were identified to the genus level, and of the 181 isolates resolved to the species level, 167 isolates (92.2%) could be assigned an accurate species identification. We also evaluated a subset of isolates for preanalytic factors that might influence MALDI-TOF MS identification. Frequent subcultures increased the number of unidentified isolates. Incubation temperatures and subcultures of the media did not alter the rate of identification. These data define the ideal bacterial preparation, identification score, and medium conditions for optimal identification of Gram-positive bacteria by use of MALDI-TOF MS.

Original languageEnglish (US)
Pages (from-to)1421-1427
Number of pages7
JournalJournal of Clinical Microbiology
Volume51
Issue number5
DOIs
StatePublished - 2013

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formic acid
Mass Spectrometry
Lasers
Gram-Positive Bacteria
Temperature

ASJC Scopus subject areas

  • Microbiology (medical)
  • Medicine(all)

Cite this

Optimizing identification of clinically relevant gram-positive organisms by use of the bruker biotyper matrix-assisted laser desorption ionization-time of flight mass spectrometry system. / TeKippe, Erin McElvania; Shuey, Sunni; Winkler, David W.; Butler, Meghan A.; Burnham, Carey Ann D.

In: Journal of Clinical Microbiology, Vol. 51, No. 5, 2013, p. 1421-1427.

Research output: Contribution to journalArticle

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