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Mechanisms of Resistance

Emergence of Plasmid-Mediated Quinolone Resistance in Escherichia coli in Europe

Hedi Mammeri, Marc Van De Loo, Laurent Poirel, Luis Martinez-Martinez, Patrice Nordmann
Hedi Mammeri
1Service de Bactériologie-Virologie, Hôpital de Bicêtre, Assistance Publique/Hôpitaux de Paris, Faculté de Médecine Paris-Sud, Université Paris XI, Le Kremlin-Bicêtre, France
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Marc Van De Loo
1Service de Bactériologie-Virologie, Hôpital de Bicêtre, Assistance Publique/Hôpitaux de Paris, Faculté de Médecine Paris-Sud, Université Paris XI, Le Kremlin-Bicêtre, France
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Laurent Poirel
1Service de Bactériologie-Virologie, Hôpital de Bicêtre, Assistance Publique/Hôpitaux de Paris, Faculté de Médecine Paris-Sud, Université Paris XI, Le Kremlin-Bicêtre, France
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Luis Martinez-Martinez
2Servicio de Microbiologia, Hospital Universitario Marquès de Valdecilla, Santander, Spain
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Patrice Nordmann
1Service de Bactériologie-Virologie, Hôpital de Bicêtre, Assistance Publique/Hôpitaux de Paris, Faculté de Médecine Paris-Sud, Université Paris XI, Le Kremlin-Bicêtre, France
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  • For correspondence: nordmann.patrice@bct.ap-hop-paris.fr
DOI: 10.1128/AAC.49.1.71-76.2005
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  • FIG. 1.
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    FIG. 1.

    Plasmid DNAs from clinical isolate E. coli Lo and E. coli transconjugant strains (A) and Southern hybridization of plasmid DNAs with the qnr-specific probe (B) and the blaVEB-1-specific probe (C). Lanes: 1, E. coli Lo; 2, E. coli J53/pQR1; 3, E. coli J53/pMG252 (used as a positive control); M, E. coli NCTC50192 (used as a negative control and a reference for plasmid sizes).

  • FIG. 2.
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    FIG. 2.

    Comparison of sul1-type integrons that contain a qnr gene (A) and the blaVEB-1-positive class 1 integron In53 (B). Shaded boxes in In53 indicate gene cassettes possessing their own promoter sequences.

  • FIG. 3.
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    FIG. 3.

    Nucleotide sequence of the promoter structure for qnr expression, as determined by the 5′ RACE experiment. The deduced amino acid sequence of Qnr is designated in single-letter code below the nucleotide sequence. The transcription orientation of the qnr gene is indicated by a horizontal arrow. The right-hand boundary of CR1 is shaded in gray. The −35 and −10 promoter sequences are boxed, as is the +1 transcription initiation site; all of these are part of the CR1 element. The vertical arrow indicates the position of the extremity of the CR1 right-hand boundary of In36 (24), in which the 67-bp sequence with a dotted underlined is lacking.

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  • TABLE 1.

    Primers used in this study

    PrimerSequenceSource or reference
    ORF513D35′-CTCACGCCCTGGCAAGGTTT-3′This study
    ORF513D55′-CTTTTGCCCTAGCTGCGGT-3′This study
    CMLA-B5′-TGGGATTTGATGTACTTTCCG-3′ 13
    CMLA-F5′-CAAAAACTTTAGTTGGCGGTAC-3′ 13
    aadB-B5′-CGCATATCGCGACCTGAAAGC-3′ 13
    aadB-F5′-GATACACAAAATTCTAGCTGCG-3′ 13
    arr-2F5′-AATTACAAGCAGGTGCAAGGA-3′ 13
    arr-2B5′-TTCAATGACGTGTAAACCACG-3′ 13
    5′CS5′-GGCATCCAAGCAGCAAG-3′ 13
    3′CS5′-AAGCAGACTTGACCTGA-3′ 13
    VEB-1A5′-CGACTTCCATTTCCCGAT GC-3′ 13
    VEB-1B5′-GGACTCTGCAACAAATACGC-3′ 13
    VEB-INV35′-GAACAGAATCAGTTCCTCCG-3′This study
    VEB-INV45′-ACGAAGAACAAATGCACAAGG-3′This study
    OXA-10CASB5′-CTTTGTTTT AGCCACCAATGATG-3′ 13
    OXA-10CASF5′-TTAGGCCTCGCCGAAGCG-3′ 13
    OrfG-B5′-GTCATTTTGAACTGCATTACC-3′This study
    IS26A5′-CTTACCAGGCGCATTTCGCC-3′This study
    AAC1-F5′-GTGAATTATTGCGGAATGCAGC 12
    Sul1A5′-CTT CGATGAGAGCCGGCGGC-3′This study
    Sul1B5′-GCAAGGCGGAAACCCGCGCC-3′This study
    QnrA5′-GGGTATGGATATTATTGATAAAG-3′ 24
    QnrB5′-CTAATCCGGCAGCACTATTA-3′ 24
    GyrA65′-CGACCTTGCGAGAGAAAT-3′ 8
    gyrA631R5′-GTTCCATCAGCCCTTCAA-3′ 8
    ParCF435′-AGCGCCTTGCGTACATGAAT-3′ 7
    ParCF9815′-GTGGTAGCGAAGAGGTGGTT-3′ 7
    PreTEM-15′-GTATCCGCTCATGAGACAATA-3′ 7
    PreTEM-25′-TCTAAAGTATATATGAGTAAACTTGGT 7
    SHV-F5′-ATGCGTTATWTTCGCCTGTGT-3′ 4
    SHV-B5′-TTAGCGTTGCCAGTGCTCG-3′ 4
    GES1A5′-ATGCGCTTCATTCACGCAC-3′ 4
    GES1B5′-CTATTTGTCCGTGCTCAGG-3′ 4
    PER-A5′-ATGAATGTCATTATAAAAGC-3′ 4
    PER-D5′-AATTTGGGCTTAGGGCAGAA-3′ 4
    GSP15′-AAGTACATCTTATGGCTGACT-3′This study
    GSP25′-ATGAAACTGCAATCCTCGAAACTG-3′This study
    GSP35′-TGGCTGAAGTCACACTGATAAAAG-3′This study
  • TABLE 2.

    MICs of antibiotics for E. coli Lo, transconjugant E. coli J53/pQR1, and E. coli J53 Az

    Antibiotic(s)MIC (μg/ml)
    E. coli LoaE. coli J53/pQR1E. coli J53
    Nalidixic acid>256324
    Ofloxacin410.12
    Ciprofloxacin10.250.01
    Moxifloxacin20.50.03
    Sparfloxacin410.01
    Rifampin>256328
    Chloramphenicol32324
    Gentamicin1680.12
    Tobramycin128640.12
    Streptomycin>256>2562
    Amikacin1680.25
    Sulfamethoxazole>512>5120.12
    Trimethoprim>512>5120.12
    Tetracycline>6411
    Amoxicillin>512>5124
    Amoxicillin-CLAb128324
    Ticarcillin>512>5124
    Ticarcillin-CLA25644
    Piperacillin128162
    Piperacillin-TZBc421
    Cephalothin1281288
    Cefoxitin422
    Cefotaxime440.06
    Ceftazidime2562560.06
    Ceftazidime + CLA0.250.250.06
    Aztreonam32320.06
    Cefepime110.01
    Imipenem0.250.250.06
    • ↵ a E. coli Lo and E. coli J53/pQR1 contained the qnr-positive plasmid that also harbored the blaVEB-1 gene.

    • ↵ b CLA, clavulanic acid at 2 μg/ml.

    • ↵ c TZB, tazobactam at 4 μg/ml.

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Emergence of Plasmid-Mediated Quinolone Resistance in Escherichia coli in Europe
Hedi Mammeri, Marc Van De Loo, Laurent Poirel, Luis Martinez-Martinez, Patrice Nordmann
Antimicrobial Agents and Chemotherapy Dec 2004, 49 (1) 71-76; DOI: 10.1128/AAC.49.1.71-76.2005

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Emergence of Plasmid-Mediated Quinolone Resistance in Escherichia coli in Europe
Hedi Mammeri, Marc Van De Loo, Laurent Poirel, Luis Martinez-Martinez, Patrice Nordmann
Antimicrobial Agents and Chemotherapy Dec 2004, 49 (1) 71-76; DOI: 10.1128/AAC.49.1.71-76.2005
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KEYWORDS

anti-infective agents
Drug Resistance, Bacterial
Escherichia coli
Escherichia coli Proteins
plasmids
quinolones

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