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Ambler Class A Extended-Spectrum β-Lactamases in Pseudomonas aeruginosa: Novel Developments and Clinical Impact

Gerhard F. Weldhagen, Laurent Poirel, Patrice Nordmann
Gerhard F. Weldhagen
1Department of Medical Microbiology, Institute of Pathology, Faculty of Health Sciences, University of Pretoria, 0001 Pretoria, South Africa
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Laurent Poirel
2Service de Bactériologie-Virologie, Hôpital de Bicêtre, Assistance Publique/Hôpitaux de Paris, Faculté de Médecine Paris-Sud, 94275 Le Kremlin-Bicêtre, France
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Patrice Nordmann
2Service de Bactériologie-Virologie, Hôpital de Bicêtre, Assistance Publique/Hôpitaux de Paris, Faculté de Médecine Paris-Sud, 94275 Le Kremlin-Bicêtre, France
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  • For correspondence: nordmann.patrice@bct.ap-hop-paris.fr
DOI: 10.1128/AAC.47.8.2385-2392.2003
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  • FIG. 1.
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    FIG. 1.

    Dendrogram obtained for class A ESBLs identified in P. aeruginosa by parsimony analysis. The alignments used for construction of the tree were carried out with the ClustalW program, followed by minor adjustments to fit the class A β-lactamase scheme (1). Branch lengths are drawn to scale and are proportional to the number of amino acid changes. The number of changes is indicated above each branch. The distance along the vertical axis has no significance.

  • FIG. 2.
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    FIG. 2.

    Double-disk synergy test performed with imipenem (IPM)- and ceftazidime (CAZ)-containing disks and a GES-1-producing P. aeruginosa isolate without (A) or with (B) cloxacillin (200 μg/ml)-containing Mueller-Hinton agar plates. No synergy was visible with a cefsulodin (CFS)-containing disk.

Tables

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  • TABLE 1.

    Ambler class A extended-spectrum β-lactamases known in P. aeruginosa

    β-LactamaseGenetic supportaCountry of first isolationYr of first isolationOther countries of isolationReference(s)
    VEB-1C, P, IFrance1998Thailand, India,b Chinab8, 17, 32, 51
    VEB-1aC, IKuwait199943
    VEB-1bC, IKuwait199943
    VEB-2C, IThailand199917
    PER-1CFrance1991Turkey, Italy, Belgium10, 12, 24, 34, 53, 54
    SHV-2aC, PFrance1995Thailand, Polandb8, 31
    SHV-5PThailand1994-1996Greeceb8
    SHV-12CThailand1994-19968
    TEM-4P, CFrance199642
    TEM-21CFrance199713
    TEM-24PFrance199825
    TEM-42PFrance199230
    GES-1C, IFrance199914
    GES-2P, ISouth Africa200044, 45
    IBC-2C, IGreece199827
    • ↵ a C, chromosomal location; P, plasmid borne; I, integron-borne.

    • ↵ b P. Nordmann, personal data.

  • TABLE 2.

    Percent amino acid identity between representatives of each type of Ambler class A ESBL identified in P. aeruginosa

    ESBL% Amino acid identity
    TEM-4SHV-2aVEB-1PER-1
    SHV-2a63
    VEB-11921
    PER-1182038
    GES-131301923
  • TABLE 3.

    Comparative kinetic parameters for ESBLs found in P. aeruginosaa

    AntibioticGES-1GES-2PER-1VEB-1TEM-4SHV-2a
    kcat (s−1)Km (μM)kcat/ Kmkcat (s−1)Km (μM)kcat/ Kmkcat (s−1)Km (μM)kcat/ KmVmaxKm (μM)Vmax/ KmVmaxKm (μM)Vmax/ KmVmaxKm (μM)Vmax/ Km
    Benzylpenicillin2.840700.44967.231.32301002.8100100NANA33171.9
    Amoxicillin13200650.725.826NANANA1106.05050NANANANANA
    AmpicillinNANANANANANANANANANANANANANANA47281.7
    Ticarcillin0.34000.70.0613.34.5NANANA8122NANANANANANA
    Piperacillin8900130.322.823NANANANANANANANANANANANA
    Cephalothin1793,400520.3311212.4462697006.0325NANANANANANA
    Cephaloridin532,000260.57.76513237222,30012533232NANA100303.2
    Cefoxitin0.93033NHNHNHNANANANANANA< 1NANANANANA
    CefuroximeNANANANANANANANANA2,00024230NANANANANANA
    Ceftazidime3802,000188ND>3,000ND703,51919.9NANANA10NANANANANA
    Cefotaxime684,600152.28902.543652664,30038314300NANA7260.2
    Cefepime2.81,8001.61.119000.6NANANANANANANANANANANANA
    CefpiromeNANANANANANANANANANANANANANANA201510.14
    Imipenem0.003450.070.0040.45NANANANANANANA<1NANANANANA
    MeropenemNHNHNHNHNHNHNANANANANANANANANANANANA
    AztreonemNHNHNHNHNHNH4344297.34005002<1NANANANANA
    • ↵ a Data were adapted from references 5, 36, 40, 41, and 45. kcat values are available only for GES-1 and GES-2. Vmax values are relative to that of benzylpenicillin and, for VEB-1, TEM-4 and SHV2a, relative to that of cephaloridine, which were set equal to 100. Abbreviations: NA, data not available; NH, not hydrolyzed (the initial rate of hydrolysis is reported to be lower than 0.001 μM−1 s−1); ND, not determinable due to Km values that are too high.

  • TABLE 4.

    MICs of β-lactams for several representative class A ESBL-producing P. aeruginosa isolatesa

    β-LactambMIC (μg/ml) for the following extended-spectrum β-lactamase (isolate denomination):
    VEB-1 (JES-1)PER-1 (RNL-1)GES-1 (695)GES-2 (GW-1)SHV2a (RP-1)TEM-4 (Stel)IBC-2 (555)
    Ticarcillin>512512>512>512>512>512>256
    Ticarcillin + CLA25625664>5126432>256
    Piperacillin1283251212825632>256
    Piperacillin + TZBNAcNA64128168>256
    Ceftazidime5121283232328>256
    Ceftazidime + CLA1284321684NA
    CefotaximeNA64NA128>512128>256
    Cefepime128NA1632NA8NA
    Imipenem320.511624>128
    Meropenem8NANA16NA1>128
    Aztrenom>256256416321632
    • ↵ a Data were adapted from references 14, 27, 30 to 32, 34, 42, and 45.

    • ↵ b CLA, clavulanic acid at a fixed concentration of 2 μg/ml; TZB, tazobactam at a fixed concentration of 4 μg/ml.

    • ↵ c NA, data not available.

  • TABLE 5.

    Primers used for detection of genes encoding class A ESBLs in P. aeruginosa

    Primer nameSequence (5′ to 3′)Gene detectedReference(s)
    VEB-1ACGACTTCCATTTCCCGATGC bla VEB 43
    VEB-1BGGACTCTGCAACAAATACGC
    PER-AATGAATGTCATTATAAAAGC bla PER 10, 45
    PER-BAATTTGGGCTTAGGGCAGAA
    GES-1AATGCGCTTCATTCACGCAC bla GES 45
    GES-1BCTATTTGTCCGTGCTCAGG
    TEM-AGAGTATTCAACATTTCCGTGTC bla TEM 45
    TEM-BTAATCAGTGAGGCACCTATCTC
    SWSHV-AAAGATCCACTATCGCCAGCAG bla SHV 31
    SWSHV-BATTCAGTTCCGTTTCCCAGCGG
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Ambler Class A Extended-Spectrum β-Lactamases in Pseudomonas aeruginosa: Novel Developments and Clinical Impact
Gerhard F. Weldhagen, Laurent Poirel, Patrice Nordmann
Antimicrobial Agents and Chemotherapy Jul 2003, 47 (8) 2385-2392; DOI: 10.1128/AAC.47.8.2385-2392.2003

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Ambler Class A Extended-Spectrum β-Lactamases in Pseudomonas aeruginosa: Novel Developments and Clinical Impact
Gerhard F. Weldhagen, Laurent Poirel, Patrice Nordmann
Antimicrobial Agents and Chemotherapy Jul 2003, 47 (8) 2385-2392; DOI: 10.1128/AAC.47.8.2385-2392.2003
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    • EPIDEMIOLOGY
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KEYWORDS

Pseudomonas Infections
Pseudomonas aeruginosa
beta-lactamases

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