Resistance to piperacillin/tazobactam in Escherichia coli resulting from extensive IS26-associated gene amplification of blaTEM-1

Katrine Hartung Hansen, Minna Rud Andreasen, Martin Schou Pedersen, Henrik Westh, Lotte Jelsbak, Kristian Schønning

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review


blaTEM-1 encodes a narrow-spectrum b-lactamase that is inhibited by b-lactamase inhibitors and commonly present in Escherichia coli. Hyperproduction of blaTEM-1 may cause resistance to penicillin/b-lactamase
inhibitor (P/BLI) combinations.

To characterize EC78, an E. coli bloodstream isolate, resistant to P/BLI combinations, which contains extensive amplification of blaTEM-1 within the chromosome.

EC78 was sequenced using Illumina and Oxford Nanopore Technology (ONT) methodology. Configuration of blaTEM-1 amplification was probed using PCR. Expression of blaTEM-1 mRNA was determined usingquantitative PCR and b-lactamase activity was determined spectrophotometrically in a nitrocefin conversion assay. Growth rate was assessed to determine fitness and stability of the gene amplification was assessed by passage in the absence of antibiotics.

Illumina sequencing of EC78 identified blaTEM-1B as the only acquired b-lactamase preceded by the WT P3 promoter and present at a copy number of 182.6 with blaTEM-1B bracketed by IS26 elements. The chromosomal location of the IS26-blaTEM-1B amplification was confirmed by ONT sequencing. Hyperproduction of blaTEM-1 was confirmed by increased transcription of blaTEM-1 and b-lactamase activity and associated with a significant fitness cost; however, the array was maintained at a relatively high copy number for 150 generations. PCR screening for blaTEM amplification of isolates resistant to P/BLI combinations identified an additional strain containing an IS26-associated amplification of a blaTEM gene.

IS26-associated amplification of blaTEM can cause resistance to P/BLI combinations. This adaptive mechanism of resistance may be overlooked if simple methods of genotypic prediction (e.g. gene presence/absence)
are used to predict antimicrobial susceptibility from sequencing data.
TidsskriftJournal of Antimicrobial Chemotherapy
Udgave nummer11
Sider (fra-til)3179-3183
Antal sider5
StatusUdgivet - 2019

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