Surveillance and Detection of Inhibitor-Resistant Beta-Lactamases in Clinical Isolates of Escherichia coli


In recent years, resistance to beta-lactam antibiotics, such as the widely-used cephalosporins and penicillins, has become a major challenge for disease therapy, particularly in common hospital-acquired infections. In the search for the mechanisms behind this increasingly prevalent form of resistance, microbiologists have identified a new type of beta-lactamase enzyme, called inhibitor-resistant TEMs (IRTs), which can withstand the effects of beta-lactamase inhibitor compounds, further reducing the arsenal of drugs available to physicians facing resistant bacteria. In this study, we examined the enzymatic and genetic basis of Escherichia coli isolates demonstrating such resistance to beta-lactam/beta-lactamase inhibitor combinations. Susceptibility trials played a major role in composing the experimental cohort for this project (n=50); each isolate was thoroughly tested to ensure that it was resistant to ampicillin-sulbactam, an inhibitor combination, but susceptible to the third-generation cephalosporin ceftazidime. Subsequently, a number of samples were subjected to assay by pulsed-field gel electrophoresis (n=18) and polymerase chain reaction (n=3) so that their genetic composition and relatedness might be known. In particular, the presence of genes coding for TEM-type beta-lactamases was investigated for each of the 3 isolates sequenced. Even though it was anticipated that the isolates would possess resistance to inhibitor combinations due to an IRT gene, this was not found to be the case. Instead, the mechanism of resistance turned out to be over-expression of a gene coding for a normal TEM enzyme. The results of these experiments have implications for ensuring successful therapy of bacterial infections and for preventing the spread of antimicrobial resistance.