Detection of different classes of carbapenemases: Adaptation and assessment of a phenotypic method applied to Enterobacteriaceae, Pseudomonas aeruginosa and Acinetobacter baumannii, and proposal of a new algorithm

J Microbiol Methods. 2018 Apr:147:26-35. doi: 10.1016/j.mimet.2018.02.013. Epub 2018 Feb 24.

Abstract

A new phenotypic method for detecting carbapenemases has been adapted (assembling of two MAST® kits, including one that contains faropenem to which a temocillin disk has been added) then assessed using 101 bacterial strains (Enterobacteriaceae with assays on Pseudomonas aeruginosa and Acinetobacter baumannii) including 62 which produce genetically identified carbapenemases. Concerning Carbapenemase-Producing Enterobacteriaceae (CPE), there is 100% sensitivity for Klebsiella pneumoniae carbapenemase (KPC, Ambler class A) and OXA-48 (Ambler class D), and 91% for metallo-beta-lactamase (MBL, Ambler class B), with a 97% sensitivity for all carbapenemases, with a specificity of 100%. The test is also efficient for detecting Pseudomonas aeruginosa carbapenemases (sensitivity between 82 and 100% and 100% specificity). The major innovation is the combined use of faropenem and temocillin for reliable detection (excellent performance with 100% sensitivity and specificity) of OXA-48. This study has led to the development of a new algorithm to detect the different classes of carbapenemases, for first-line diagnosis, by combining this modified MAST® test with immunochromatographic methods and molecular biology techniques.

Keywords: Algorithm; Carbapenemases; Enterobacteriaceae; Faropenem; OXA-48; Temocillin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acinetobacter baumannii / enzymology*
  • Algorithms*
  • Bacterial Proteins / classification
  • Bacterial Proteins / isolation & purification*
  • Bacteriological Techniques / methods*
  • Enterobacteriaceae / enzymology*
  • Microbial Sensitivity Tests / methods*
  • Molecular Biology / methods
  • Penicillins / pharmacology
  • Phenotype
  • Pseudomonas aeruginosa / enzymology*
  • Reagent Kits, Diagnostic*
  • Sensitivity and Specificity
  • beta-Lactamases / classification
  • beta-Lactamases / isolation & purification*
  • beta-Lactams / pharmacology

Substances

  • Bacterial Proteins
  • Penicillins
  • Reagent Kits, Diagnostic
  • beta-Lactams
  • temocillin
  • beta-Lactamases
  • carbapenemase
  • fropenem