Related Experiment Video
Updated: Jun 26, 2026
Isolation and Identification of Waterborne Antibiotic-Resistant Bacteria and Molecular Characterization of their Antibiotic Resistance Genes
Published on: March 3, 2023
A simple disc diffusion method for detecting AmpC and extended-spectrum beta-lactamases in clinical isolates of
Helen Derbyshire1, Gemma Kay, Katie Evans
1Department of Microbiology, Royal Hallamshire Hospital, Glossop Road, Sheffield, UK.
Background:
We sought to determine whether extended-spectrum beta-lactamases (ESBLs) and AmpC beta-lactamases (derepressed and inducible), alone and in combination, could be detected in unidentified members of the Enterobacteriaceae using a simple, overnight disc diffusion test.
Methods:
The genetic basis of antibiotic resistance in cephalosporin-resistant wild-type (n = 140) and culture collection (n = 140) isolates of Enterobacteriaceae was determined using PCR. A scheme for detecting these resistance mechanisms phenotypically was devised using five antibiotic discs: cefpodoxime +/- clavulanate; cefepime +/- clavulanate and cefoxitin.
Results And Conclusions:
AmpC beta-lactamases (derepressed and inducible) and ESBLs, alone and in combination, could reliably be detected using a disc diffusion method. ESBLs alone could be detected on the basis of a difference of >5 mm between cefpodoxime/clavulanate and cefpodoxime (10 microg) discs. ESBLs, in the presence of AmpC beta-lactamases, could be detected using a difference of >5 mm between cefepime/clavulanate and cefepime (30 microg) discs. AmpC beta-lactamases could be detected using a difference of >14 mm between cefepime/clavulanate and cefpodoxime/clavulanate discs. Inducible AmpC beta-lactamases could be discerned by observing the blunting of the cefpodoxime or cefpodoxime/clavulanate zones in proximity to cefoxitin (30 microg) discs.
