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Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

Editor-in-Chief: Plebani, Mario

Ed. by Gillery, Philippe / Greaves, Ronda / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Payne, Deborah A. / Schlattmann, Peter


IMPACT FACTOR 2018: 3.638

CiteScore 2018: 2.44

SCImago Journal Rank (SJR) 2018: 1.191
Source Normalized Impact per Paper (SNIP) 2018: 1.205

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1437-4331
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Volume 57, Issue 8

Issues

Rapid susceptibility testing of multi-drug resistant Escherichia coli and Klebsiella by glucose metabolization monitoring

Maximilian Kittel
  • Institute for Clinical Chemistry, Medical Faculty Mannheim of Heidelberg University, Mannheim, Germany
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/ Peter Findeisen
  • Institute for Clinical Chemistry, Medical Faculty Mannheim of Heidelberg University, Mannheim, Germany
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/ Beniam Ghebremedhin
  • Institute for Medical Laboratory Diagnostics, Center for Clinical and Translational Research, HELIOS Klinikum Wuppertal, Witten/Herdecke University, Witten, Germany
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/ Thomas Miethke
  • Institute for Medical Microbiology and Hygiene, Medical Faculty Mannheim of Heidelberg University, Mannheim, Germany
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/ Alexander Grundt
  • Institute for Clinical Chemistry, Medical Faculty Mannheim of Heidelberg University, Mannheim, Germany
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/ Parviz Ahmad-Nejad
  • Institute for Medical Laboratory Diagnostics, Center for Clinical and Translational Research, HELIOS Klinikum Wuppertal, Witten/Herdecke University, Witten, Germany
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/ Michael Neumaier
  • Corresponding author
  • Institute for Clinical Chemistry, Medical Faculty Mannheim of Heidelberg University, Mannheim, Germany, Phone: +49 – 621 383 2222, Fax: +49 – 621 383 3819
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Published Online: 2019-02-14 | DOI: https://doi.org/10.1515/cclm-2018-1178

Abstract

Background

The increasing number of multi-drug resistant (MDR) bacteria provides enormous challenges for choosing an appropriate antibiotic therapy in the early phase of sepsis. While bacterial identification has been greatly accelerated by the introduction of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), the antibiotic susceptibility testing (AST) remains time-consuming. Here, we present a rapid susceptibility testing method for testing Gram-negative bacteria, exemplarily validated for Escherichia coli and Klebsiella spp.

Methods

Gram-negative isolates (E. coli and Klebsiella spp.) were either taken as single colonies from agar plates (n=136) or directly extracted and identified from positive blood cultures (n=42) using MALDI-TOF MS. Bacteria were incubated in glucose-supplemented Luria broths (LBs) each containing one antibiotic (ceftazidime, piperacillin, imipenem and ciprofloxacin), routinely used to classify Gram-negative bacteria in Germany. To determine susceptibility the dynamics of glucose utilization in bacterial suspensions were quantitatively measured in the presence or absence of antibiotics designated liquid-AST (L-AST).

Results

The L-AST can be run on clinical-chemistry analyzers and integrated into laboratory routines. It yields critical resistance information within 90–150 min downstream of a MS-based identification. The results showed a high concordance with routine susceptibility testing, with less than 1% very major errors (VME) and 3.51% major errors (ME) for 178 assessed isolates. Analysis of turnaround time (TAT) for 42 clinical samples indicated that L-AST results could be obtained 34 h earlier than the routine results.

Conclusions

As exemplified for E. coli and Klebsiella spp., L-AST provides substantial acceleration of susceptibility testing following MALDI-TOF MS identification. The assay is a simple and low-cost method that can be integrated into clinical laboratory to allow for 24/7 AST. This approach could improve antibiotic therapy.

This article offers supplementary material which is provided at the end of the article.

Keywords: antibiotic susceptibility testing; glucose; multi-drug resistant (MDR); rapid antibiotic susceptibility testing

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About the article

Received: 2018-11-02

Accepted: 2019-01-16

Published Online: 2019-02-14

Published in Print: 2019-07-26


Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

Research funding: None declared.

Employment or leadership: None declared.

Honorarium: None declared.

Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.


Citation Information: Clinical Chemistry and Laboratory Medicine (CCLM), Volume 57, Issue 8, Pages 1271–1279, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/cclm-2018-1178.

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