An optimized work-flow to reduce time-to-detection of carbapenemase-producing Enterobacteriaceae (CPE) using direct testing from rectal swabs

Bioengineered. 2017 May 4;8(3):217-224. doi: 10.1080/21655979.2016.1222335. Epub 2016 Aug 17.

Abstract

Rapid detection of patients with carbapenemase-producing Enterobacteriaceae (CPE) is essential for the prevention of nosocomial cross-transmission, allocation of isolation facilities and to protect patient safety. Here, we aimed to design a new laboratory work-flow, utilizing existing laboratory resources, in order to reduce time-to-diagnosis of CPE. A review of the current CPE testing processes and of the literature was performed to identify a real-time commercial polymerase chain reaction (PCR) assay that could facilitate batch testing of CPE clinical specimens, with adequate CPE gene coverage. Stool specimens (210) were collected; CPE-positive inpatients (n = 10) and anonymized community stool specimens (n = 200). Rectal swabs (eSwab™) were inoculated from collected stool specimens and a manual DNA extraction method (QIAamp® DNA Stool Mini Kit) was employed. Extracted DNA was then processed on the Check-Direct CPE® assay. The three step process of making the eSwab™, extracting DNA manually and running the Check-Direct CPE® assay, took <5 min, 1 h 30 min and 1 h 50 min, respectively. It was time efficient with a result available in under 4 h, comparing favourably with the existing method of CPE screening; average time-to-diagnosis of 48/72 h. Utilizing this CPE work-flow would allow a 'same-day' result. Antimicrobial susceptibility testing results, as is current practice, would remain a 'next-day' result. In conclusion, the Check-Direct CPE® assay was easily integrated into a local laboratory work-flow and could facilitate a large volume of CPE screening specimens in a single batch, making it cost-effective and convenient for daily CPE testing.

Keywords: carbapenemase-producing; check-direct CPE®; eSwab™; Enterobacteriaceae; polymerase chain reaction.

MeSH terms

  • Algorithms
  • Bacterial Proteins / biosynthesis
  • Bacterial Typing Techniques / methods*
  • Enterobacteriaceae / enzymology
  • Enterobacteriaceae / isolation & purification*
  • Enterobacteriaceae Infections / diagnosis*
  • Enterobacteriaceae Infections / microbiology*
  • Feces / microbiology*
  • Humans
  • Rectum / microbiology
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Specimen Handling / methods
  • Workflow*
  • Workload
  • beta-Lactamases / biosynthesis

Substances

  • Bacterial Proteins
  • beta-Lactamases
  • carbapenemase

Grants and funding

This study was funded by a research grant awarded by the Irish Society of Clinical Microbiologists, which is supported by Pfizer Ireland.