Phenotypic and molecular detection methods for carbapenemase-producing organisms and their clinical significance at two Scottish tertiary care hospitals

J Med Microbiol. 2019 Apr;68(4):560-565. doi: 10.1099/jmm.0.000931. Epub 2019 Mar 14.

Abstract

Purpose: This study evaluated in-house PCR testing for local identification of bacteria carrying the major carbapenemase genes (blaOXA-48-like, blaVIM, blaNDM, blaKPC and blaIMP).

Methodology: Carbapenemase-producing organisms (CPOs) isolated from patients managed in two tertiary care hospitals in Scotland from September 2014-January 2017 were investigated. A combination of chromogenic screening agar (ChromID CARBA SMART), a carbapenem hydrolysis test (Rapidec Carba NP) and in-house real-time PCR for the blaOXA-48-like, blaVIM, blaNDM, blaKPC and blaIMP genes were utilized. All isolates were sent to the AMRHAI reference unit for confirmatory testing.

Results: During the 29-month study period 39 CPO were isolated from 34 patients. The average turnaround time for a workflow involving phenotypic and molecular testing was 4.2 days. PCR had a sensitivity and specificity of 100 %. The most common carbapenemase genes were blaOXA-48-like (31%), blaVIM (23%) and blaNDM (20%). Resistance to antimicrobials other than beta-lactams was common; the most active agents were colistin, amikacin and fosfomycin. Twenty-seven patients were considered to be colonized (although CPO detection influenced empiric antimicrobials in five) and a CPO was implicated in infection in seven patients (bacteraemia in immunocompromised patients, n=2; surgical site infections, n=2; osteomyelitis in a patient with diabetes mellitus, n=1; and urinary tract infections, n=2). All patients survived infection.

Conclusion: In a lowincidence setting we demonstrate the efficacy of a combined local laboratory workflow for rapid detection of CPOs, incorporating phenotypic and molecular testing. In 7/34 patients the CPO was implicated as a pathogen and detection influenced antimicrobial decision-making in five colonized patients.

Keywords: antimicrobial resistance; carbapenem resistance; carbapenem-producing Enterobacteriaceae; carbapenemase-producing organisms; molecular diagnostics.

Publication types

  • Evaluation Study

MeSH terms

  • Aged
  • Anti-Bacterial Agents / pharmacology
  • Bacteremia / diagnosis
  • Bacterial Proteins / biosynthesis*
  • Carbapenems / pharmacology
  • Enterobacteriaceae / enzymology
  • Enterobacteriaceae / genetics*
  • Enterobacteriaceae Infections / diagnosis
  • Enterobacteriaceae Infections / microbiology*
  • Female
  • Humans
  • Hydrolysis
  • Male
  • Microbial Sensitivity Tests
  • Middle Aged
  • Molecular Diagnostic Techniques
  • Phenotype
  • Real-Time Polymerase Chain Reaction
  • Scotland
  • Sensitivity and Specificity
  • Tertiary Care Centers
  • beta-Lactamases / biosynthesis*

Substances

  • Anti-Bacterial Agents
  • Bacterial Proteins
  • Carbapenems
  • beta-Lactamases
  • carbapenemase