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

Resumé

Background:
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.

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

Methods:
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.

Results:
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.

Conclusions:
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.
OriginalsprogEngelsk
TidsskriftJournal of Antimicrobial Chemotherapy
Vol/bind74
Udgave nummer11
Sider (fra-til)3179-3183
ISSN0305-7453
DOI
StatusUdgivet - 2019

Citer dette

Hansen, Katrine Hartung ; Andreasen, Minna Rud ; Pedersen, Martin Schou ; Westh, Henrik ; Jelsbak, Lotte ; Schønning, Kristian. / Resistance to piperacillin/tazobactam in Escherichia coli resulting from extensive IS26-associated gene amplification of blaTEM-1. I: Journal of Antimicrobial Chemotherapy. 2019 ; Bind 74, Nr. 11. s. 3179-3183.
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title = "Resistance to piperacillin/tazobactam in Escherichia coli resulting from extensive IS26-associated gene amplification of blaTEM-1",
abstract = "Background: 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-lactamaseinhibitor (P/BLI) combinations.Objectives: To characterize EC78, an E. coli bloodstream isolate, resistant to P/BLI combinations, which contains extensive amplification of blaTEM-1 within the chromosome.Methods: 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.Results: 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.Conclusions: 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.",
author = "Hansen, {Katrine Hartung} and Andreasen, {Minna Rud} and Pedersen, {Martin Schou} and Henrik Westh and Lotte Jelsbak and Kristian Sch{\o}nning",
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doi = "10.1093/jac/dkz349",
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pages = "3179--3183",
journal = "Journal of Antimicrobial Chemotherapy",
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Resistance to piperacillin/tazobactam in Escherichia coli resulting from extensive IS26-associated gene amplification of blaTEM-1. / Hansen, Katrine Hartung ; Andreasen, Minna Rud; Pedersen, Martin Schou; Westh, Henrik ; Jelsbak, Lotte; Schønning, Kristian.

I: Journal of Antimicrobial Chemotherapy, Bind 74, Nr. 11, 2019, s. 3179-3183.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Resistance to piperacillin/tazobactam in Escherichia coli resulting from extensive IS26-associated gene amplification of blaTEM-1

AU - Hansen, Katrine Hartung

AU - Andreasen, Minna Rud

AU - Pedersen, Martin Schou

AU - Westh, Henrik

AU - Jelsbak, Lotte

AU - Schønning, Kristian

PY - 2019

Y1 - 2019

N2 - Background: 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-lactamaseinhibitor (P/BLI) combinations.Objectives: To characterize EC78, an E. coli bloodstream isolate, resistant to P/BLI combinations, which contains extensive amplification of blaTEM-1 within the chromosome.Methods: 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.Results: 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.Conclusions: 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.

AB - Background: 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-lactamaseinhibitor (P/BLI) combinations.Objectives: To characterize EC78, an E. coli bloodstream isolate, resistant to P/BLI combinations, which contains extensive amplification of blaTEM-1 within the chromosome.Methods: 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.Results: 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.Conclusions: 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.

U2 - 10.1093/jac/dkz349

DO - 10.1093/jac/dkz349

M3 - Journal article

VL - 74

SP - 3179

EP - 3183

JO - Journal of Antimicrobial Chemotherapy

JF - Journal of Antimicrobial Chemotherapy

SN - 0305-7453

IS - 11

ER -