Abstract
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.
Keywords:
ESRI; Escherichia coli; beta-lactamase; beta-lactamase inhibitor; piperacillin; resistance; tazobactam.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Anti-Bacterial Agents / pharmacology
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Bacteriological Techniques
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Drug Resistance, Multiple, Bacterial* / drug effects
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Escherichia coli / genetics
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Escherichia coli / isolation & purification*
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Escherichia coli Infections / diagnosis*
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Escherichia coli Infections / microbiology
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Humans
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Lactams / pharmacology
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Microbial Sensitivity Tests
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Piperacillin, Tazobactam Drug Combination / pharmacology*
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beta-Lactamase Inhibitors / pharmacology*
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beta-Lactamases / genetics
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beta-Lactams / pharmacology
Substances
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Anti-Bacterial Agents
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Lactams
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beta-Lactamase Inhibitors
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beta-Lactams
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Piperacillin, Tazobactam Drug Combination
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beta-Lactamases