Rapid detection of antibiotic resistance based on mass spectrometry and stable isotopes

  • J. S. Jung
  • T. Eberl
  • K. Sparbier
  • C. Lange
  • M. Kostrzewa
  • S. Schubert
  • A. Wieser
Article
First Online:
Received:
Accepted:

Abstract

With the emergence and growing complexity of bacterial drug resistance, rapid and reliable susceptibility testing has become a topical issue. Therefore, new technologies that assist in predicting the effectiveness of empiric antibiotic therapy are of great interest. Although the use of matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS) for the rapid detection of antibiotic resistance is an attractive option, the current methods for MALDI-TOF MS susceptibility testing are restricted to very limited conditions. Here, we describe a technique that may allow for rapid susceptibility testing to an extent that is comparable to phenotypic methods. The test was based on a stable isotope labelling by amino acids in cell culture (SILAC)-like approach. This technique was used to visualise the growth of bacteria in the presence of an antibiotic. Pseudomonas aeruginosa was chosen as the model organism, and strains were incubated in normal medium, medium supplemented with 13C6-15 N2-labelled lysine and medium supplemented with labelled lysine and antibiotic. Peak shifts occurring due to the incorporation of the labelled amino acids were detected by MALDI-TOF MS. Three antibiotics with different mechanisms of action, meropenem, tobramycin and ciprofloxacin, were tested. A semi-automated algorithm was created to enable rapid and unbiased data evaluation. With the proposed test, a clear distinction between resistant and susceptible isolates was possible for all three antibiotics. The application of SILAC technology for the detection of antibiotic resistance may contribute to accelerated and reliable susceptibility testing.

Keywords

Minimum Inhibitory Concentration Meropenem Peak Shift EUCAST Label Amino Acid 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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Notes

Acknowledgements

This study was supported by a grant from the Bayerische Forschungsstiftung (Forschungsverbund “ForBIMed—Biomarker in der Infektionsmedizin”) to S.S. and M.K.

Conflict of interest

Katrin Sparbier, Christoph Lange and Markus Kostrzewa are employed at the mass spectrometry company Bruker Daltonik GmbH. The other authors declare that they have no conflicts of interest.

Supplementary material

10096_2013_2031_Fig4_ESM.gif (15 kb)
Fig S1

P. aeruginosa ATCC 27853 and one P. aeruginosa clinical isolate from a broth culture at different growth phases were inoculated into DMEM medium containing labelled lysine and non-labelled lysine. Incorporation of the labelled amino acids in the lag phase (90 min), early growth phase (180 min), exponential growth phase (300 min) and stationary phase (660 min) was measured in triplicate. Ratios of the intensities of labelled to non-labelled peaks are displayed in a box plot diagram. No obvious correlation between the growth phase and the incorporation of the labelled amino acid was detected. (GIF 14 kb)

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High-resolution image (EPS 3645 kb)
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Fig S2

Susceptibility testing was performed with ciprofloxacin and four representative strains of P. aeruginosa (two susceptible and two resistant). The isolates were grown on Columbia blood and MacConkey agar. To allow for comparison with the normal inter-assay variability, the results of two different days are displayed. No differences related to the medium of the starter culture were observed. (GIF 16 kb)

10096_2013_2031_MOESM2_ESM.eps (3.6 mb)
High-resolution image (EPS 3654 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • J. S. Jung
    • 1
  • T. Eberl
    • 1
  • K. Sparbier
    • 2
  • C. Lange
    • 2
  • M. Kostrzewa
    • 2
  • S. Schubert
    • 1
  • A. Wieser
    • 1
  1. 1.Max von Pettenkofer-Institut für Hygiene und Medizinische MikrobiologieMunichGermany
  2. 2.Bruker Daltonik GmbHBremenGermany

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