ena-STUDY-Erasmus University Medical Center-12-04-2021-13:12:06:513-824
LC-MS/MS analysis demonstrates a decrease in porins and increase in CMY-2-like beta-lactamases in E. coli exposed to meropenem
While ESBL and AmpC beta-lactamases barely degrade carbapenems, they are able to bind them and prevent them from interacting with penicillin binding proteins thereby preventing their effect. When these beta-lactamases are expressed at a high level and combined with a decreased influx of carbapenems due to a decrease in membrane permeability, Enterobacterales can become resistant to carbapenems. In this study we developed a LC-MS/MS assay for the detection of the E. coli porins OmpC and OmpF, it's chromosomal AmpC beta-lactamase and the plasmid-mediated CMY-2 beta-lactamase. Subsequently, we cultured CMY-2 positive E. coli isolates in the presence of meropenem and analyzed mutants that showed increased resistance to meropenem using our developed assay and western blot. In all five selected strains, a decrease in OmpC and/or OmpF was the first event towards an increase in meropenem minimum inhibitory concentrations (MICs). Subsequently, in four of the five isolate series, MICs increased further after an increase in CMY-2-like production.
While ESBL and AmpC beta-lactamases barely degrade carbapenems, they are able to bind them and prevent them from interacting with penicillin binding proteins thereby preventing their effect. When these beta-lactamases are expressed at a high level and combined with a decreased influx of carbapenems due to a decrease in membrane permeability, Enterobacterales can become resistant to carbapenems. In this study we developed a LC-MS/MS assay for the detection of the E. coli porins OmpC and OmpF, it's chromosomal AmpC beta-lactamase and the plasmid-mediated CMY-2 beta-lactamase. Subsequently, we cultured CMY-2 positive E. coli isolates in the presence of meropenem and analyzed mutants that showed increased resistance to meropenem using our developed assay and western blot. In all five selected strains, a decrease in OmpC and/or OmpF was the first event towards an increase in meropenem minimum inhibitory concentrations (MICs). Subsequently, in four of the five isolate series, MICs increased further after an increase in CMY-2-like production.