Evaluation of Piperacillin-Tazobactam ETEST for the Detection of OXA-1 Resistance Mechanism among Escherichia coli and Klebsiella pneumoniae

Globally, piperacillin-tazobactam resistance among Escherichia coli and Klebsiella pneumoniae is driven by OXA-1 beta-lactamases. Expression of bla(OXA-1) yields piperacillin-tazobactam MICs of 8 to 16 mu g/mL, which straddle the susceptible/susceptible-dose dependent breakpoint set by the Clinical and Laboratory Standards Institute in 2022. Variability of the reference broth microdilution method (BMD) was evaluated by manufacturing BMD panels using 2 brands of piperacillin, 2 brands of tazobactam and 2 brands of cation-adjusted Mueller-Hinton broth. In addition, ETEST, which harbors an intermediate dilution of 12 mu g/mL was evaluated for the ability to differentiate isolates with and without bla(OXA-1). A collection of 200 E. coli and K. pneumoniae, of which 82 harbored a bla(OXA-1) gene, were tested. BMD variability was on average 1.3-fold, within the accepted 2-fold variability of MICs. However, categorical agreement (CA) between BMD reads was 74.0% for all isolates and 63.4% for those with a bla(OXA-1) gene and 81.3% for those without bla(OXA-1) detected (P = 0.004, Pearson's Chi Square). ETEST overall CA with the BMD mode was 68.0% and essential agreement (EA) was 80.5%. For isolates with bla(OXA-1), CA was 50.0% and EA was 69.5%, versus 80.5% and 88.1%, respectively, for isolates without bla(OXA-1) (P < 0.0001 for both comparisons). All ETEST errors were major errors (false resistance) compared to BMD mode. However, the negative predictive value of the ETEST for the presence of bla(OXA-1) was 94.1%, compared to only 74.2% negative predictive value for BMD. Clinicians and microbiologists should be aware of the challenges associated with testing piperacillin-tazobactam in regions where bla(OXA-1) is prevalent. Globally, piperacillin-tazobactam resistance among Escherichia coli and Klebsiella pneumoniae is driven by OXA-1 beta-lactamases. Expression of bla(OXA-1) yields piperacillin-tazobactam MICs of 8 to 16 mu g/mL, which straddle the susceptible/susceptible-dose dependent breakpoint set by the Clinical and Laboratory Standards Institute in 2022.

Automated summary

Globally, piperacillin-tazobactam resistance among Escherichia coli and Klebsiella pneumoniae is mainly driven by OXA-1 beta-lactamases. The variability of the reference broth microdilution method (BMD) and ETEST in differentiating isolates with and without bla(OXA-1) gene was evaluated. ETEST showed lower accuracy and higher false resistance compared to BMD.