Semirapid Detection of Piperacillin/Tazobactam Resistance and Extended-Spectrum Resistance to β-Lactams/β-Lactamase Inhibitors in Clinical Isolates of Escherichia coli.

Piperacillin/tazobactam (TZP) is a β-lactam/β-lactamase inhibitor (BL/BLI) recommended for the empirical treatment of severe infections. The excessive and indiscriminate use of TZP has promoted the emergence of TZP-resistant Escherichia coli isolates. Recently, we demonstrated that TZP may contribute to the development of extended-spectrum resistance to BL/BLI (ESRI) in E. coli isolates that are TZP susceptible but have low-level resistance to BL/BLI (resistance to amoxicillin/clavulanic acid [AMC] and/or ampicillin/sulbactam [SAM]). This raises the need for the development of rapid detection systems. Therefore, the objective of this study was to design and validate a method able to detect TZP resistance and ESRI in E. coli. A colorimetric assay based on β-lactam ring hydrolysis by β-lactamases was designed (ESRI test). A total of 114 E. coli isolates from bloodstream and intra-abdominal sources, characterized according to their susceptibility profiles to BL/BLI, were used. Detection of the three most frequent β-lactamases involved in BL/BLI resistance (blaTEM, blaOXA-1, and blaSHV) was performed by PCR. The ESRI test was able to detect all the TZP-intermediate/-resistant isolates, as well as all the TZP-susceptible isolates with a capacity for ESRI development. Their median times to results were 5 and 30 min, respectively. All the isolates without resistance to BL/BLI displayed a negative result in the ESRI test. blaTEM was the most frequent β-lactamase gene detected, follow by blaSHV and blaOXA-1. These results demonstrate the efficacy of the ESRI test, showing great clinical potential which could lead to reductions in health costs, ineffective treatments, and inappropriate use of BL/BLI. IMPORTANCE TZP is a BL/BLI recommended for the empirical treatment of severe infections. The excessive use of TZP has promoted the emergence of TZP-resistant Escherichia coli isolates. We recently reported that TZP may contribute to the development of ESRI in E. coli isolates that are TZP susceptible but have low-level resistance to BL/BLI. This raises the need for the development of rapid detection systems. Here, we demonstrated that the ESRI test was able to detect the TZP-intermediate or -resistant isolates and the TZP-susceptible isolates with the capacity for ESRI development. All the isolates without BL/BLI resistance were negative for the ESRI test and did not harbor β-lactamase genes. For ESRI developers and TZP-intermediate or -resistant isolates, blaTEM was the most frequent β-lactamase gene detected, follow by blaSHV and blaOXA-1. The sensitivity, specificity, and positive and negative predictive values were all 100%. These data demonstrate the efficacy of the ESRI test and show that it has great clinical potential.