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An Improved Medium for Colistin Susceptibility Testing.

Konrad Gwozdzinski1,2, Saina Azarderakhsh1,2, Can Imirzalioglu1,2

  • 1Institute of Medical Microbiology, Justus Liebig University, Giessen, Germany.

Journal of Clinical Microbiology
|March 9, 2018
PubMed
Summary
This summary is machine-generated.

A new calcium-enhanced Mueller-Hinton (CE-MH) medium accurately detects colistin resistance in bacteria, including those with the mcr-1 gene. This improved medium simplifies susceptibility testing for laboratories, even in resource-limited settings.

Keywords:
calcium ionscolistin susceptibility testingmcr-1 resistance

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Area of Science:

  • Clinical Microbiology
  • Antimicrobial Resistance
  • Bacterial Genetics

Background:

  • The plasmid-mediated colistin resistance gene, mcr-1, confers resistance to colistin, a critical last-line antibiotic against multidrug-resistant Gram-negative bacteria.
  • Current colistin susceptibility testing methods using cation-adjusted Mueller-Hinton (CA-MH) medium misclassify approximately 15% of mcr-1-positive bacteria as susceptible.
  • Accurate colistin susceptibility testing is crucial for guiding treatment of infections caused by multidrug-resistant Gram-negative bacteria.

Purpose of the Study:

  • To develop and evaluate an improved calcium-enhanced Mueller-Hinton (CE-MH) medium for reliable colistin susceptibility testing.
  • To assess the performance of CE-MH medium in differentiating between colistin-resistant and colistin-susceptible bacterial isolates, including those with mcr-1.
  • To determine the utility of CE-MH medium in routine diagnostic laboratories.

Main Methods:

  • Colistin susceptibility testing was performed on 50 mcr-1-positive Enterobacteriaceae isolates (E. coli, K. pneumoniae), 7 intrinsically polymyxin-resistant species, and 32 mcr-1-negative isolates.
  • Testing was conducted using both broth microdilution (BMD) and strip-based formats with both CA-MH and the novel CE-MH media.
  • Isogenic IncX4 plasmid lacking mcr-1 was used for data validation.

Main Results:

  • The CE-MH medium demonstrated clear separation between resistant and susceptible isolates in both BMD and gradient diffusion assays.
  • This improved performance was observed for mcr-1-containing Enterobacteriaceae, as well as isolates with intrinsic or acquired colistin resistance.
  • CE-MH medium is simple to prepare and overcomes limitations of current testing methods.

Conclusions:

  • The calcium-enhanced Mueller-Hinton (CE-MH) medium provides a simple and reliable method for accurate colistin susceptibility testing.
  • CE-MH medium effectively detects resistance mediated by the mcr-1 gene and other resistance mechanisms.
  • This novel medium is easily implementable in routine diagnostic laboratories, including those in resource-poor settings.