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MALDI-TOF MS applied to indirect carbapenemase detection: a validated procedure to clearly distinguish between carbapenemase-positive and carbapenemase-negative bacterial strains

Analytical and Bioanalytical Chemistry volume 405, pages 5259–5266 (2013)Cite this article

Abstract

Laboratory identification of carbapenemase-producing clinical isolates is crucial to limit the spread of the bacteria. In this study, we shall first develop the matrix-assisted laser desorption ionization–time-of-flight mass spectrometry (MALDI-TOF MS) assay in automatic identification of carbapenemase producers. A total of 143 well-characterized isolates were studied. After an incubation of bacteria with meropenem trihydrate, the mixture was centrifuged and the supernatant analyzed by MALDI-TOF MS. A genetic algorithm model with ClinProTools software was built using spectra of 43 carbapenemase-positive isolates and 40 carbapenemase-negative isolates after 2 h of incubation. This model was externally validated using 60 test isolates. All spectra of supernatants of the carbapenemase-negative isolates showed peak profiles comparable to that of pure meropenem (m/z 384.159, 406.140, and 428.122 of its two sodium salt variants) regardless of the incubation time tested. For the carbapenemase-positive isolates, the specific peak for meropenem at m/z 384.159 disappeared during the incubation time, two products of meropenem degradation were identified with m/z 358.18 (the decarboxylated product) and 380.161 (sodium salt of the decarboxylated product), and other degradation products were observed (native molecule with disrupted amide bond with m/z 402.169, three sodium salt variants with m/z 424.151, 446.133, and 468.115). Sixty test isolates were 100 % correctly classified as carbapenemase positive and carbapenemase negative with the genetic algorithm model. MALDI-TOF MS coupled with ClinProTools is capable of rapidly, accurately, and automatically identifying carbapenemase producers.

The average spectra of the carbapenemase-positive (red) and carbapenemasenegative isolates (green) were shown. Nine peaks differentiating the two classes are highlighted by arrows. x axis, mass per charge [m/z (in daltons)]; y axis, intensity(arbitrary units [arb.u.]).

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Acknowledgments

We thank Dr. Dong Li (Bruker Daltonics, China) for his assistance during the study. This work was supported by the Research Foundation of Beijing Tongren Hospital Affiliated to Capital Medical University (no. 2012-YJJ-010).

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Laboratory Medicine, Beijing Tongren Hospital, No. 1. Dongjiaominxiang Road, Dongcheng District, Beijing, 100730, China

    Lijun Wang, Wenjun Sui, Mei Wang & Xinxin Lu

  2. Department of Laboratory Medicine, Xi’an No. 1 Hospital, Xi’an City 029, Shanxi Province, China

    Chao Han

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  1. Lijun Wang
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  2. Chao Han
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  3. Wenjun Sui
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  4. Mei Wang
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Correspondence to Xinxin Lu.

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Wang, L., Han, C., Sui, W. et al. MALDI-TOF MS applied to indirect carbapenemase detection: a validated procedure to clearly distinguish between carbapenemase-positive and carbapenemase-negative bacterial strains. Anal Bioanal Chem 405, 5259–5266 (2013). https://doi.org/10.1007/s00216-013-6913-2

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  • DOI: https://doi.org/10.1007/s00216-013-6913-2

Keywords

  • MALDI
  • Carbapenem
  • Carbapenemases
  • Resistance
  • Classification