Enhanced detection of carbapenemase-producing Enterobacteriaceae by an optimized phenol red assay

Diagn Microbiol Infect Dis. 2018 Jan;90(1):11-17. doi: 10.1016/j.diagmicrobio.2017.09.005. Epub 2017 Sep 18.

Abstract

Screening for the detection of carbapenemase-producing bacteria still encounters issues related to workflow, limit of detection, or qualitative interpretation. We developed a spectrophotometry-based version of the Carba NP phenol red assay (Nordmann et al., 2012) in a microtiter plate format, compatible with low bacterial cell counts. We were able to detect highly active carbapenemases such as KPC and IMP in 30min. A wider range of carbapenemases including OXA-48 were detected using higher inocula, still being competitive compared with currently available phenol red assays. Validation experiments of our test with a panel of 81 Enterobacteriaceae showed good performance with 93% of sensitivity and 92% of specificity. The compatibility of our routine-friendly protocol with automation offers great perspectives for high throughput screening in outbreak situations and/or in big laboratories.

Keywords: Assay; Bacteria; Carbapenemase; Detection; Phenol-red; Spectrophotometry.

MeSH terms

  • Bacterial Proteins / metabolism*
  • Bacteriological Techniques / methods*
  • Biological Assay / methods*
  • Carbapenem-Resistant Enterobacteriaceae / drug effects*
  • Carbapenem-Resistant Enterobacteriaceae / enzymology
  • Carbapenem-Resistant Enterobacteriaceae / metabolism
  • Drug Resistance, Bacterial / physiology
  • Enterobacter cloacae / drug effects
  • Enterobacter cloacae / genetics
  • Enterobacteriaceae Infections / drug therapy
  • Enterobacteriaceae Infections / microbiology
  • Escherichia coli / drug effects
  • Escherichia coli / genetics
  • Escherichia coli Proteins / genetics
  • Humans
  • Klebsiella pneumoniae / drug effects
  • Klebsiella pneumoniae / genetics
  • Phenolsulfonphthalein / chemistry*
  • beta-Lactamases / genetics
  • beta-Lactamases / metabolism*

Substances

  • Bacterial Proteins
  • Escherichia coli Proteins
  • beta-Lactamases
  • beta-lactamase OXA-48, E coli
  • carbapenemase
  • Phenolsulfonphthalein