Utility of a novel multiplex TaqMan PCR assay for metallo-β-lactamase genes plus other TaqMan assays in detecting genes encoding serine carbapenemases and clinically significant extended-spectrum β-lactamases

Rosemary Swayne, Matthew Ellington*, Martin Curran, Neil Woodford, Sani H. Aliyu

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

32 Citations (Scopus)

Abstract

Prompt detection of infections caused by Enterobacteriaceae that produce therapeutically important β-lactamases [metallo-β-lactamases (MBLs), serine carbapenemases, acquired AmpC and CTX-M extended-spectrum β-lactamases (ESBLs)] is crucial for infection prevention and control and surveillance purposes, and, more contentiously, also for effective patient management. A novel TaqMan PCR assay was developed to detect genes encoding IMP, VIM, NDM, SPM, SIM and GIM MBLs. Published PCR assays for acquired genes encoding CTX-M ESBLs and AmpC β-lactamases were updated and adapted to the TaqMan format, respectively. A published TaqMan assay for serine carbapenemase genes was used. Assay specificity was tested using a panel of 59 isolates with known acquired genes from the four different β-lactamase groupings. The four TaqMan assays correctly identified the most clinically relevant acquired β-lactamase genes in the panel of 59 resistant Enterobacteriaceae, which included 3 VIM-, 7 NDM- and 12 IMP-producers. Consecutive, non-duplicate isolates of Enterobacteriaceae from 965 urinary and 343 blood cultures during 2010 were then screened for β-lactamase genes using these TaqMan assays. Amongst the urinary and blood culture isolates tested, 69 CTX-M-producers and 21 acquired AmpC β-lactamase-producers were identified; the CTX-M rate amongst blood culture isolates (9.3%) broadly reflects the UK national average. During the study period, one Klebsiella pneumoniae isolate producing an NDM carbapenemase was identified from a wound sample. The assays developed and/or used will enable the future surveillance and the rapid detection and appropriate early treatment of infections caused by Gram-negative bacteria producing clinically important β-lactamases, including carbapenemases.

Original languageEnglish
Pages (from-to)352-356
Number of pages5
JournalInternational Journal of Antimicrobial Agents
Volume42
Issue number4
DOIs
Publication statusPublished - Oct 2013

Bibliographical note

Funding Information:
Funding : This study was supported by Public Health England (formerly the Health Protection Agency) and Cambridge University Hospitals NHS Foundation Trust .

Funding Information:
Competing interests : MJE, NW and MDC work for Public Health England and are influenced by its views on antimicrobial resistance and antimicrobial use; MDC has received research funding from the Health Protection Agency (HPA); MJE has received research funding from the HPA, National Institute for Health Research (NIHR) and AstraZeneca; NW has received research funding from the HPA, NIHR, AstraZeneca, the UK government's Department of Health and Merck; SHA has received sponsorship to attend conferences from Gilead Sciences, Schering Plough and Wyeth. RS declares no competing interests.

Keywords

  • Carbapenemases
  • Cephalosporinases
  • Enterobacteriaceae
  • Extended-spectrum β-lactamases
  • Metallo-β-lactamases

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