Detection of extended-spectrum β-lactamase and carbapenemase activity in Gram-negative bacilli using liquid chromatography - tandem mass spectrometry

Serafim, Vlad, Shah, Ajit J. ORCID: https://orcid.org/0000-0002-2350-6384, Licker, Monica ORCID: https://orcid.org/0000-0002-9245-5883, Horhat, Florin George ORCID: https://orcid.org/0000-0001-6133-0204, Vulpie, Silvana, Musuroi, Corina and Muntean, Delia ORCID: https://orcid.org/0000-0001-9100-4530 (2020) Detection of extended-spectrum β-lactamase and carbapenemase activity in Gram-negative bacilli using liquid chromatography - tandem mass spectrometry. Infection and Drug Resistance, 13 . pp. 4021-4029. ISSN 1178-6973 [Article] (doi:10.2147/IDR.S267160)

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Abstract

PURPOSE: Several mass spectrometry-based methods for antimicrobial sensitivity testing have been described in recent years. They offer an alternative to commercially available testing systems which were considered to have disadvantages in terms of cost- and time-efficiency. The aim of this study was to develop a LC-MS/MS-based antibiotic hydrolysis assay for evaluating antimicrobial resistance of Gram-negative bacteria.

METHODS: Four species of Gram-negative bacilli (Klebsiella pneumoniae, Escherichia coli, Providencia stuartii and Acinetobacter baumannii) were tested against six antibiotics from three different classes: ampicillin, meropenem, imipenem, ceftazidime, ceftriaxone and cefepime. Bacterial suspensions from each species were incubated with a mixture of the six antibiotics. Any remaining antibiotic following incubation were measured using LC-MS/MS. The results were interpreted using measurements obtained for an E. coli strain sensitive to all antibiotics and expressed as percentage of hydrolyzed antibiotic. These were subsequently compared to commercially-available system for the bacteria identification and susceptibility testing.

RESULTS: Overall, LC-MS/MS assay and commercial antimicrobial susceptibility platform results showed good agreement in terms of an organism being resistant/sensitive to an antibiotic. The time required to complete the LC-MS/MS-based hydrolysis test was under 5 h, significantly shorter that commercially available susceptibility testing platforms.

CONCLUSION: By using a sensitive strain for results interpretation and simultaneous use of multiple antibiotics, the proposed protocol offers improved robustness and multiplexing over previously described methods for antibiotic sensitivity testing. Nevertheless, further research is needed before routine assimilation of the method, especially for strains with intermediate resistance.

Item Type: Article
Keywords (uncontrolled): Gram-negative, LC-MS/MS, antimicrobial, mass spectrometry, resistance, β-lactamases
Research Areas: A. > School of Science and Technology > Natural Sciences
Item ID: 30941
Notes on copyright: © 2020 Serafim et al. This work is published and licensed by Dove Medical Press Limited. The full terms of this license are available at https://www.dovepress.com/terms.php and incorporate the Creative Commons Attribution – Non Commercial (unported, v3.0) License (http://creativecommons.org/licenses/by-nc/3.0/). By accessing the work you hereby accept the Terms. Non-commercial uses of the work are permitted without any further permission from Dove Medical Press Limited, provided the work is properly attributed. For permission for commercial use of this work, please see paragraphs 4.2 and 5 of our Terms (https://www.dovepress.com/terms.php).
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Depositing User: Ajit Shah
Date Deposited: 11 Sep 2020 14:09
Last Modified: 07 Feb 2021 18:30
URI: https://eprints.mdx.ac.uk/id/eprint/30941

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