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

  • Maximilian Kittel , Peter Findeisen , Beniam Ghebremedhin , Thomas Miethke , Alexander Grundt , Parviz Ahmad-Nejad und Michael Neumaier EMAIL logo
Veröffentlicht/Copyright: 14. Februar 2019
Clinical Chemistry and Laboratory Medicine (CCLM)
Aus der Zeitschrift Clinical Chemistry and Laboratory Medicine (CCLM) Band 57 Heft 8

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.

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

Acknowledgments

We gratefully acknowledge the technical assistance of Cornelia Mathies-Schwarz, Laura Voss, Corinna Mosbach, and Ruth Leiblein in the development and validation of L-AST.

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

  2. Research funding: None declared.

  3. Employment or leadership: None declared.

  4. Honorarium: None declared.

  5. 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.

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Supplementary Material

The online version of this article offers supplementary material (https://doi.org/10.1515/cclm-2018-1178).

Received: 2018-11-02
Accepted: 2019-01-16
Published Online: 2019-02-14
Published in Print: 2019-07-26

©2019 Walter de Gruyter GmbH, Berlin/Boston

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https://doi.org/10.1515/cclm-2018-1178
Schlagwörter für diesen Artikel
antibiotic susceptibility testing; glucose; multi-drug resistant (MDR); rapid antibiotic susceptibility testing

Artikel in diesem Heft

  1. Frontmatter
  2. Editorials
  3. CCLM Award for the Most Cited Paper
  4. Folate and vitamin B12 assays after recalibration to the WHO International Standard 03/178: making the interpretation as simple as possible, but not simpler
  5. Reviews
  6. Blood contamination in salivary diagnostics: current methods and their limitations
  7. Central adrenal insufficiency: open issues regarding diagnosis and glucocorticoid treatment
  8. Genetics and Molecular Diagnostics
  9. Measuring the chronology of the translational process of molecular genetic discoveries
  10. Development and interlaboratory evaluation of a NIST Reference Material RM 8366 for EGFR and MET gene copy number measurements
  11. General Clinical Chemistry and Laboratory Medicine
  12. Post-translational modification-derived products are associated with frailty status in elderly subjects
  13. Urine chloride self-measurement to monitor sodium chloride intake in patients with chronic kidney disease
  14. Estimated urinary osmolality based on combined urinalysis parameters: a critical evaluation
  15. Measurement of S100B protein: evaluation of a new prototype on a bioMérieux Vidas® 3 analyzer
  16. Measuring thyroglobulin in patients with thyroglobulin autoantibodies: evaluation of the clinical impact of BRAHMS Kryptor® Tg-minirecovery test in a large series of patients with differentiated thyroid carcinoma
  17. Human chorionic gonadotropin suspected heterophile interference investigations in immunoassays: a recommended approach
  18. Certified reference material against PR3 ANCA IgG autoantibodies. From development to certification
  19. Diagnostic accuracy of a fully automated multiplex celiac disease antibody panel for serum and plasma
  20. Fasting serum bile acids concentration is associated with insulin resistance independently of diabetes status
  21. Hematology and Coagulation
  22. The association between activated protein C ratio and Factor V Leiden are gender-dependent
  23. Reference Values and Biological Variations
  24. Determination of sigma score based on biological variation for haemostasis assays: fit-for-purpose for daily practice?
  25. Calcitonin measurement in pediatrics: reference ranges are gender-dependent, validation in medullary thyroid cancer and thyroid diseases
  26. Cancer Diagnostics
  27. Uncovering the clinical impact of kallikrein-related peptidase 5 (KLK5) mRNA expression in the colorectal adenoma-carcinoma sequence
  28. Cardiovascular Diseases
  29. Performance of a novel high sensitivity cardiac troponin I assay in asymptomatic hemodialysis patients – evidence for sex-specific differences
  30. Infectious Diseases
  31. Rapid susceptibility testing of multi-drug resistant Escherichia coli and Klebsiella by glucose metabolization monitoring
  32. Letters to the Editor
  33. Vitamin B12 and folate levels in a healthy population: establishing reference intervals
  34. Reference values of a new serum folate assay traceable to the WHO International Standard
  35. Serum protein electrophoresis and complement deficiencies: a veteran but very versatile test in clinical laboratories
  36. Introduction of a novel ELISA assay for serum AMH determination
  37. Bone alkaline phosphatase on the IDS-iSYS automated analyser; cross-reactivity with intestinal ALP
  38. Evaluation of the MULTISURE HIV Rapid Test in a Korean population with low human immunodeficiency virus prevalence
  39. Vancomycin immunoassay: does the Advia Centaur XPT underestimate the exposure of patients? A method comparison study
  40. Hb Hunan and Hb Hengyang: Two unexpected discoveries during HbA1c measurements
  41. Analytical performance of a CE-marked immunoassay to quantify phosphorylated neurofilament heavy chains
  42. Implementation of an automated method for direct quantification of urinary ammonium
  43. Congress Abstracts
  44. Proceedings of ACBI 2018 41ST Annual Conference Association of Clinical Biochemists in Ireland
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