Multiplex PCR to detect pAmpC β-lactamases among enterobacteriaceae at a tertiary care laboratory in Mumbai, India

Microbiology (Reading). 2019 Feb;165(2):246-250. doi: 10.1099/mic.0.000748. Epub 2018 Dec 13.

Abstract

Drug-resistance due to AmpC β-lactamases represents a growing problem worldwide. In this study, a previously collected sample of 108 cefoxitin-resistant clinical isolates was assessed for AmpC β-lactamase production through routine phenotypic testing and double-disc cefoxitin/cloxcallin (DD-CC), cefoxitin/phenylboronic acid (CDT-PBA) and AmpC disc tests. The same isolates were characterized by a novel multiplex polymerase chain reaction molecular assay to detect the presence of blaACT, blaDHA, blaCIT, blaFOX, blaMIR and blaMOX. By phenotypic analysis, 56%, 55% and 48 % were detected as being AmpC β-lactamase producers by the CDT-PBA, DD-CC and AmpC disc tests, respectively. By molecular analysis, 57 % were determined to be AmpC β-lactamase producers, including 34 % blaFOX, 8 % blaCIT and 1.6 % blaDHAas mono-AmpC producers. The production of multiple AmpC molecular types was common, including 30 % with both blaCIT+FOX and 1.6 % each of blaCIT+DHA, blaACT+MIR, blaACT+FOX, blaACT+DHA and blaMIR+FOX. Molecular characterization of AmpC would help detect the prevalence of AmpC β-lactamase producers, facilitate proper patient management and implement infection control practices.

Keywords: AmpC disk test; Cefoxtin-Cloxcallin; Molecular characterization; ampC β-lactamases; cefoxitin-resistance.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Bacterial Agents / pharmacology
  • Bacterial Proteins / biosynthesis
  • Bacterial Proteins / genetics*
  • Cefoxitin / pharmacology
  • Enterobacteriaceae / drug effects
  • Enterobacteriaceae / genetics*
  • Enterobacteriaceae / isolation & purification
  • Enterobacteriaceae / metabolism
  • Enterobacteriaceae Infections / epidemiology
  • Enterobacteriaceae Infections / microbiology
  • Genotype
  • Humans
  • India / epidemiology
  • Microbial Sensitivity Tests
  • Multiplex Polymerase Chain Reaction / methods*
  • Phenotype
  • Sensitivity and Specificity
  • Tertiary Healthcare
  • beta-Lactam Resistance / drug effects
  • beta-Lactam Resistance / genetics
  • beta-Lactamases / biosynthesis
  • beta-Lactamases / genetics*

Substances

  • Anti-Bacterial Agents
  • Bacterial Proteins
  • Cefoxitin
  • AmpC beta-lactamases
  • beta-Lactamases