Rectal screening for carbapenemase-producing Enterobacteriaceae: a proposed workflow

J Glob Antimicrob Resist. 2020 Jun:21:86-90. doi: 10.1016/j.jgar.2019.10.012. Epub 2019 Oct 19.

Abstract

Objectives: Active screening is a crucial element for the prevention of carbapenemase-producing Enterobacteriaceae (CPE) transmission in healthcare settings. Here we propose a culture-based protocol for rectal swab CPE screening that combines CPE detection with identification of the carbapenemase type.

Methods: The workflow integrates an automatic digital analysis of selective chromogenic media (WASPLab®; Copan), with subsequent rapid tests for the confirmation of carbapenemase production [i.e. detection of Klebsiella pneumoniae carbapenemase (KPC)-specific peak by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF/MS) or a multiplex immunochromatographic assay identifying the five commonest carbapenemase types]. To evaluate the performance of this protocol in depth, data for 21 162 rectal swabs submitted for CPE screening to the Microbiology Unit of S. Orsola-Malpighi Hospital (Bologna, Italy) were analysed.

Results: Considering its ability to correctly segregate plates with/without Enterobacteriaceae, WASPLab Image Analysis Software showed globally a sensitivity and specificity of 100% and 79.4%, respectively. Of the plates with bacterial growth (n = 901), 693 (76.9%) were found to be positive for CPE by MALDI-TOF/MS (KPC-specific peak for K. pneumoniae) or by immunochromatographic assay. Only 2.8% (16/570) of KPC-positive K. pneumoniae strains were missed by the specific MALDI-TOF/MS algorithm, being detected by the immunochromatographic assay. The mean turnaround time needed from sample arrival to the final report ranged between 18 and 24 h, representing a significant time saving compared with manual reading.

Conclusion: This workflow proved to be fast and reliable, being particularly suitable for areas endemic for KPC-producing K. pneumoniae and for high-throughput laboratories.

Keywords: CPE; Carbapenemase; Klebsiella pneumoniae; MALDI-TOF/MS; Rectal screening; WASPLab.

MeSH terms

  • Automation, Laboratory
  • Bacterial Proteins / metabolism
  • Bacteriological Techniques / methods*
  • Carbapenem-Resistant Enterobacteriaceae / growth & development*
  • Carbapenem-Resistant Enterobacteriaceae / isolation & purification
  • Carbapenem-Resistant Enterobacteriaceae / metabolism
  • Chromatography, Affinity
  • Enterobacteriaceae Infections / diagnosis*
  • High-Throughput Screening Assays
  • Humans
  • Italy
  • Rectum / microbiology*
  • Sensitivity and Specificity
  • Software
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Workflow
  • beta-Lactamases / metabolism

Substances

  • Bacterial Proteins
  • beta-Lactamases
  • carbapenemase