The performance of a resazurin chromogenic agar plate with a combined disc method for rapid screening of extended-spectrum-β-lactamases, AmpC β-lactamases and co-β-lactamases in Enterobacteriaceae

Microbiol Immunol. 2017 Aug;61(8):297-304. doi: 10.1111/1348-0421.12499.

Abstract

A promising means of rapid screening of extended-spectrum-β-lactamase (ESBL), AmpC β-lactamase, and co-production of ESBL and AmpC that combines resazurin chromogenic agar (RCA) with a combined disc method is here reported. Cefpodoxime (CPD) discs with and without clavulanic acid (CA), cloxacillin (CX) and CA+CX were evaluated against 86 molecularly confirmed β-lactamase-producing Enterobacteriaceae, including 15 ESBLs, 32 AmpCs, nine co-producers of ESBL and AmpC and 30 carbapenemase producers. The CA and CX synergy test successfully detected all ESBL producers (100% sensitivity and 98.6% specificity) and all AmpC producers (100% sensitivity and 96.36% specificity). This assay also performed well in screening for co-existence of ESBL and AmpC (88.89% sensitivity and 100% specificity). The RCA assay is simple and inexpensive and provides results within 7 hr. It can be performed in any microbiological laboratory, in particular, in geographic regions in which ESBL, AmpC or co-β-lactamase-producing Enterobacteriaceae are endemic.

Keywords: Beta-lactamases; Enterobacteriaceae; cefpodoxime combined disc; resazurin chromogenic agar.

MeSH terms

  • Anti-Bacterial Agents / pharmacology*
  • Bacterial Proteins / metabolism*
  • Cefpodoxime
  • Ceftizoxime / analogs & derivatives
  • Ceftizoxime / pharmacology
  • Clavulanic Acid / pharmacology
  • Cloxacillin / pharmacology
  • Disk Diffusion Antimicrobial Tests / methods*
  • Enterobacteriaceae / drug effects*
  • Enterobacteriaceae / isolation & purification
  • Enterobacteriaceae / metabolism
  • Humans
  • Indicators and Reagents / chemistry*
  • Oxazines / chemistry*
  • Xanthenes / chemistry*
  • beta-Lactamases / metabolism*

Substances

  • Anti-Bacterial Agents
  • Bacterial Proteins
  • Indicators and Reagents
  • Oxazines
  • Xanthenes
  • resazurin
  • Clavulanic Acid
  • Ceftizoxime
  • AmpC beta-lactamases
  • beta-Lactamases
  • carbapenemase
  • Cloxacillin