Plasmid-mediated complementation of gyrA and gyrB in fluoroquinolone-resistant Bacteroides fragilis

M. L. Peterson; J. C. Rotschafer; L. J.V. Piddock

doi:10.1093/jac/dkg368

Plasmid-mediated complementation of gyrA and gyrB in fluoroquinolone-resistant Bacteroides fragilis

M. L. Peterson, J. C. Rotschafer, L. J.V. Piddock

Experimental and Clinical Pharmacology

Research output: Contribution to journal › Article › peer-review

12 Scopus citations

Abstract

Objectives: To identify whether mutations in gyrA and gyrB confer fluoroquinolone resistance in Bacteroides fragilis. Methods: Eight fluoroquinolone-resistant (FQ^R) strains were complemented with plasmid-mediated B. fragilis wild-type gyrA (pMP1) and gyrB (pMP2), and MICs determined. Sequence analysis of the gyrA and gyrB quinolone resistance determining region (QRDR) was performed for all strains. Results: MICs of fluoroquinolones were two- to 32-fold higher than wild-type for all mutants. Five mutants had a substitution in GyrA (Ser-82→Phe), one mutant had a substitution in GyrA (Asp-81 →Gly), one mutant had a substitution in GyrB (Glu-478→Lys), and one resistant strain did not contain mutations in the QRDR of gyrA or gyrB. Following complementation with pMP1 or pMP2, the MICs of fluoroquinolones were reduced two- to 32-fold for the mutants. Conclusion: These studies verify that substitutions in GyrA and GyrB confer resistance in B. fragilis. Other mechanisms are also responsible for resistance since not all resistant strains fully complemented to the wild-type phenotype.

Original language	English (US)
Pages (from-to)	481-484
Number of pages	4
Journal	Journal of Antimicrobial Chemotherapy
Volume	52
Issue number	3
DOIs	https://doi.org/10.1093/jac/dkg368
State	Published - Sep 1 2003

Bibliographical note

Funding Information:
This study was funded by HealthPartners Research Foundation (Minneapolis, MN, USA) and the British Society for Antimicrobial Chemotherapy (BSAC) (Birmingham, UK). The authors would also like to thank Dr Abigail Salyers and Nadja Shoemaker, Department of Microbiology, University of Illinois, Urbana, IL, USA, for their assistance and contribution of several bacterial strains containing necessary plasmids, including pLYL01. In addition, M.L.P. is grateful to Dr Patrick Schlievert for his continued support throughout this work.

Keywords

Anaerobe
Antibiotic resistance
Quinolone

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10.1093/jac/dkg368

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title = "Plasmid-mediated complementation of gyrA and gyrB in fluoroquinolone-resistant Bacteroides fragilis",

abstract = "Objectives: To identify whether mutations in gyrA and gyrB confer fluoroquinolone resistance in Bacteroides fragilis. Methods: Eight fluoroquinolone-resistant (FQR) strains were complemented with plasmid-mediated B. fragilis wild-type gyrA (pMP1) and gyrB (pMP2), and MICs determined. Sequence analysis of the gyrA and gyrB quinolone resistance determining region (QRDR) was performed for all strains. Results: MICs of fluoroquinolones were two- to 32-fold higher than wild-type for all mutants. Five mutants had a substitution in GyrA (Ser-82→Phe), one mutant had a substitution in GyrA (Asp-81 →Gly), one mutant had a substitution in GyrB (Glu-478→Lys), and one resistant strain did not contain mutations in the QRDR of gyrA or gyrB. Following complementation with pMP1 or pMP2, the MICs of fluoroquinolones were reduced two- to 32-fold for the mutants. Conclusion: These studies verify that substitutions in GyrA and GyrB confer resistance in B. fragilis. Other mechanisms are also responsible for resistance since not all resistant strains fully complemented to the wild-type phenotype.",

keywords = "Anaerobe, Antibiotic resistance, Quinolone",

author = "Peterson, {M. L.} and Rotschafer, {J. C.} and Piddock, {L. J.V.}",

note = "Funding Information: This study was funded by HealthPartners Research Foundation (Minneapolis, MN, USA) and the British Society for Antimicrobial Chemotherapy (BSAC) (Birmingham, UK). The authors would also like to thank Dr Abigail Salyers and Nadja Shoemaker, Department of Microbiology, University of Illinois, Urbana, IL, USA, for their assistance and contribution of several bacterial strains containing necessary plasmids, including pLYL01. In addition, M.L.P. is grateful to Dr Patrick Schlievert for his continued support throughout this work.",

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doi = "10.1093/jac/dkg368",

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T1 - Plasmid-mediated complementation of gyrA and gyrB in fluoroquinolone-resistant Bacteroides fragilis

AU - Peterson, M. L.

AU - Rotschafer, J. C.

AU - Piddock, L. J.V.

N1 - Funding Information: This study was funded by HealthPartners Research Foundation (Minneapolis, MN, USA) and the British Society for Antimicrobial Chemotherapy (BSAC) (Birmingham, UK). The authors would also like to thank Dr Abigail Salyers and Nadja Shoemaker, Department of Microbiology, University of Illinois, Urbana, IL, USA, for their assistance and contribution of several bacterial strains containing necessary plasmids, including pLYL01. In addition, M.L.P. is grateful to Dr Patrick Schlievert for his continued support throughout this work.

PY - 2003/9/1

Y1 - 2003/9/1

N2 - Objectives: To identify whether mutations in gyrA and gyrB confer fluoroquinolone resistance in Bacteroides fragilis. Methods: Eight fluoroquinolone-resistant (FQR) strains were complemented with plasmid-mediated B. fragilis wild-type gyrA (pMP1) and gyrB (pMP2), and MICs determined. Sequence analysis of the gyrA and gyrB quinolone resistance determining region (QRDR) was performed for all strains. Results: MICs of fluoroquinolones were two- to 32-fold higher than wild-type for all mutants. Five mutants had a substitution in GyrA (Ser-82→Phe), one mutant had a substitution in GyrA (Asp-81 →Gly), one mutant had a substitution in GyrB (Glu-478→Lys), and one resistant strain did not contain mutations in the QRDR of gyrA or gyrB. Following complementation with pMP1 or pMP2, the MICs of fluoroquinolones were reduced two- to 32-fold for the mutants. Conclusion: These studies verify that substitutions in GyrA and GyrB confer resistance in B. fragilis. Other mechanisms are also responsible for resistance since not all resistant strains fully complemented to the wild-type phenotype.

AB - Objectives: To identify whether mutations in gyrA and gyrB confer fluoroquinolone resistance in Bacteroides fragilis. Methods: Eight fluoroquinolone-resistant (FQR) strains were complemented with plasmid-mediated B. fragilis wild-type gyrA (pMP1) and gyrB (pMP2), and MICs determined. Sequence analysis of the gyrA and gyrB quinolone resistance determining region (QRDR) was performed for all strains. Results: MICs of fluoroquinolones were two- to 32-fold higher than wild-type for all mutants. Five mutants had a substitution in GyrA (Ser-82→Phe), one mutant had a substitution in GyrA (Asp-81 →Gly), one mutant had a substitution in GyrB (Glu-478→Lys), and one resistant strain did not contain mutations in the QRDR of gyrA or gyrB. Following complementation with pMP1 or pMP2, the MICs of fluoroquinolones were reduced two- to 32-fold for the mutants. Conclusion: These studies verify that substitutions in GyrA and GyrB confer resistance in B. fragilis. Other mechanisms are also responsible for resistance since not all resistant strains fully complemented to the wild-type phenotype.

KW - Anaerobe

KW - Antibiotic resistance

KW - Quinolone

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JO - Journal of Antimicrobial Chemotherapy

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ER -

Plasmid-mediated complementation of gyrA and gyrB in fluoroquinolone-resistant Bacteroides fragilis

Abstract

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