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Bacteriology

Phenotypic Detection of Carbapenem-Susceptible Metallo-β-Lactamase-Producing Gram-Negative Bacilli in the Clinical Laboratory

Clare Franklin, Lisa Liolios, Anton Y. Peleg
Clare Franklin
Microbiology Unit, Alfred Pathology Service, Alfred Hospital, Commercial Road, Melbourne, Victoria 3004, Australia
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Lisa Liolios
Microbiology Unit, Alfred Pathology Service, Alfred Hospital, Commercial Road, Melbourne, Victoria 3004, Australia
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Anton Y. Peleg
Microbiology Unit, Alfred Pathology Service, Alfred Hospital, Commercial Road, Melbourne, Victoria 3004, Australia
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  • For correspondence: apeleg@bidmc.harvard.edu
DOI: 10.1128/JCM.00879-06
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  • FIG. 1.
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    FIG. 1.

    The phenotypic appearance of an MBL-producing Serratia marcescens isolate carrying the blaIMP-4 gene. (A) Combined-disk test, using two imipenem (10 μg) disks, one with 292 μg EDTA, showing an increase in zone inhibition of >4 mm around the disk with EDTA. (B) Double-disk synergy test, using an IPM (10 μg) disk placed 20 mm (center to center) from a blank filter disk containing 292 μg EDTA. (C) Aztreonam (30 μg) disk with a >30-mm zone of inhibition.

  • FIG. 2.
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    FIG. 2.

    Increase (in millimeters) in zone of inhibition around the imipenem-EDTA disk compared with the imipenem disk alone for 52 MBL-negative (MBL -ve) and 54 MBL-positive (MBL +ve) clinical isolates.

  • FIG. 3.
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    FIG. 3.

    Inhibition zone diameters (in millimeters) around the aztreonam disk (30 μg) for the 84 metallo-β-lactamase-carrying isolates. Asterisks indicate Enterobacteriaceae isolates (n = 8) found to produce an AmpC-type or extended-spectrum β-lactamase.

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  • TABLE 1.

    Results of the phenotypic method in the detection of MBL-producing gram-negative bacilli

    OrganismNo. of isolatesNo. (%) of isolates positive on phenotypic plateMBL gene type
    MBL-positive isolates84
        Serratia marcescens3737 (100)IMP-4
        P. aeruginosa1919 (100)IMP-4
        P. aeruginosa11 (100)VIM-4
        Pseudomonas stutzeri11 (100)VIM-2
        Klebsiella pneumoniae1010 (100)IMP-4
        Klebsiella oxytoca11 (100)IMP-4
        Escherichia coli66 (100)IMP-4
        Enterobacter cloacae66 (100)IMP-4
        Citrobacter koseri11 (100)IMP-4
        Acinetobacter junii11 (100)IMP-4
        Acinetobacter baumannii11 (100)IMP-1
    MBL-negative isolates52
        P. aeruginosa151 (7)NDa
        Pseudomonas mendocina10 (0)ND
        A. baumannii100 (0)ND
        E. cloacae50 (0)ND
        S. marcescens40 (0)ND
        E. coli40 (0)ND
        K. pneumoniae30 (0)ND
        K. oxytoca30 (0)ND
        Citrobacter braakii10 (0)ND
        Achromobacter xylosoxidans30 (0)ND
        Burkholderia cepacia20 (0)ND
        Ralstonia pickettii10 (0)ND
    • ↵ a ND, not detected.

  • TABLE 2.

    Summary of results for the combined-disk test and the double-disk synergy test in the detection of MBL-producing gram-negative bacilli

    Phenotypic methodNo. of PCR-confirmed MBL-carrying organisms (n = 84)No. of PCR-confirmed non-MBL-carrying organisms (n = 52)Sensitivity (%)Specificity (%)
    Combined-disk test84 (100%)1 (2%)10098
    Double-disk synergy test66 (79%)1 (2%)7998
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Phenotypic Detection of Carbapenem-Susceptible Metallo-β-Lactamase-Producing Gram-Negative Bacilli in the Clinical Laboratory
Clare Franklin, Lisa Liolios, Anton Y. Peleg
Journal of Clinical Microbiology Sep 2006, 44 (9) 3139-3144; DOI: 10.1128/JCM.00879-06

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Phenotypic Detection of Carbapenem-Susceptible Metallo-β-Lactamase-Producing Gram-Negative Bacilli in the Clinical Laboratory
Clare Franklin, Lisa Liolios, Anton Y. Peleg
Journal of Clinical Microbiology Sep 2006, 44 (9) 3139-3144; DOI: 10.1128/JCM.00879-06
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KEYWORDS

Anti-Bacterial Agents
carbapenems
Gram-negative bacteria
Gram-Negative Bacterial Infections

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