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Rapid LC–MS/MS detection of different carbapenemase types in carbapenemase-producing Enterobacterales

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European Journal of Clinical Microbiology & Infectious Diseases Aims and scope Submit manuscript

Abstract

Klebsiella pneumoniae carbapenemase (KPC)-2, metallo-beta-lactamases (MBL), and oxacillinase (OXA)-48-like carbapenemases are considered the most important carbapenemases. In vitro studies have demonstrated that the carbapenemase activity of KPC-2 and MBL can be inhibited by 3-aminophenylboronic acid and ethylenediaminetetraacetic acid (EDTA), respectively. Understanding the carbapenemase types expressed in carbapenem-resistant Enterobacterales (CRE) is of great significance to clinical therapies. Liquid chromatography-coupled tandem mass spectrometry (LC–MS/MS) is fast, stable, and specific; and is considered the gold standard method for measuring small molecules. In this study, we developed carbapenemase inhibition tests combined with LC–MS/MS to rapidly identify carbapenemase types. A total of 295 clinical isolates were examined, including 212 KPC-2 producers, 29 MBL producers, 15 OXA-48-like producers, 3 KPC-2 + OXA-232 producers, and 36 carbapenem-sensitive strains. We used LC–MS/MS to determine the carbapenemase types by measuring the ratio of the hydrolyzed meropenem peak areas in the presence and absence of different inhibitors. The sensitivity and specificity of LC–MS/MS in detecting single KPC-2 producers were 97.64% and 100.00%, respectively, and 96.55% and 100.00% for MBL producers, respectively. In addition, the sensitivity and specificity of LC–MS/MS in detecting single OXA-48-like producers were both 100.00% when extending incubation time up to 2.5 h. LC–MS/MS showed excellent agreement in detecting carbapenemase types using the modified carbapenem inactivation (mCIM)/EDTA-carbapenem inactivation assay (eCIM) (kappa = 0.93 for serine carbapenemases and kappa = 0.98 for MBL carbapenemases). In this study, LC–MS/MS demonstrated excellent detection of different carbapenemase types, showing potential reliability in future clinical applications.

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Data availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

We thank Ms. Wanjun Yu for her excellent and professional assistance in preparing this manuscript. We appreciate Professor Fupin Hu for the generous gift of OXA-48-like producing strains. This work was supported by Grants from Pudong New Area Construction of key disciplines of the Health and Family Planning Commission (grant numbers: PW2019E-2).

Funding

This work was supported by Grants from Pudong New Area Construction of key disciplines of the Health and Family Planning Commission (grant numbers: PW2019E-2).

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Author notes

  1. Gen Li, Zhihan Ye and Nianzhen Chen contributed equally to this work.

Authors and Affiliations

  1. Department of Clinical Laboratory, Shanghai East Hospital, School of Medicine, Tongji University, 150 Ji Mo Road, Shanghai, 200120, People’s Republic of China

    Gen Li, Zhihan Ye, Wenyan Zhang, Nianzhen Chen, Yangqin Ye, Yuchao Wang, Fei Wu, Keli Wang & Lieying Fan

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  1. Gen Li
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  2. Zhihan Ye
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  9. Lieying Fan
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Contributions

Gen Li and Lieying Fan designed the experiments; Gen Li, Zhihan Ye, Wenyan Zhang, Nianzhen Chen, Yangqin Ye, Yuchao Wang, Fei Wu, and Keli Wang performed the experiments; Gen Li, Zhihan Ye, and Wenyan Zhang analyzed the data; Lieying Fan contributed reagents/materials/analysis tools; Gen Li and Lieying Fan wrote the manuscript.

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Correspondence to Lieying Fan.

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Li, G., Ye, Z., Zhang, W. et al. Rapid LC–MS/MS detection of different carbapenemase types in carbapenemase-producing Enterobacterales. Eur J Clin Microbiol Infect Dis 41, 815–825 (2022). https://doi.org/10.1007/s10096-022-04440-5

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  • DOI: https://doi.org/10.1007/s10096-022-04440-5

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