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Journal of Clinical Microbiology, July 2005, p. 3129-3135, Vol. 43, No. 7
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.7.3129-3135.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Detection of Pseudomonas aeruginosa Producing Metallo-ß-Lactamases in a Large Centralized Laboratory

Johann D. D. Pitout,1,2* Daniel B. Gregson,1,2,3 Laurent Poirel,4 Jo-Ann McClure,1 Phillip Le,1 and Deirdre L. Church1,2,3

Division of Microbiology, Calgary Laboratory Services,1 Departments of Pathology and Laboratory Medicine,2 Medicine, University of Calgary, Calgary, Alberta, Canada,3 Service de Bactériologie-Virologie, Hôpital de Bicêtre, Assistance Publique/Hôpitaux de Paris, Faculté de Médecine Paris-Sud, 94275 Le Kremlin-Bicêtre, France4

Received 6 January 2005/ Returned for modification 9 March 2005/ Accepted 28 March 2005

Metallo-ß-lactamases (MBLs) have been increasingly recognized from clinical isolates worldwide, but the laboratory detection of these strains is not well defined. We report a study that developed an EDTA disk screen test and a molecular diagnostic assay for the detection of MBL-producing Pseudomonas aeruginosa. Using NCCLS disk methodology, inhibition zone diameters were determined in tests with imipenem (IPM) and meropenem (MEM) disks alone and in combination with 930 µg of EDTA. This test was compared with the MBL Etest. The duplex PCR assay showed 100% sensitivity and specificity for detecting MBL-producing control strains. Of the 241 clinical strains of IPM-nonsusceptible P. aeruginosa from the Calgary Health Region isolated from 2002 to 2004, 110/241 (46%) were MBL positive using phenotypic methods while 107/241 (45%) were PCR positive for MBL genes: 103/241 (43%) for blaVIM and 4/241 (2%) for blaIMP. The EDTA disk screen test using MEM showed 100% sensitivity and 97% specificity for detecting MBLs in control and clinical strains. The EDTA disk screen test is simple to perform and to interpret and can easily be introduced into the workflow of a clinical laboratory. We recommend that all IPM-nonsusceptible P. aeruginosa isolates be routinely screened for MBL production using the EDTA disk screen test and that PCR confirmation be performed at a regional laboratory.

* Corresponding author. Mailing address: Calgary Laboratory Services, #9, 3535 Research Road NW, Calgary, Alberta, Canada T2L 2K8. Phone: (403) 770 3309. Fax: (403) 770 3347. E-mail: johann.pitout{at}cls.ab.ca.

Journal of Clinical Microbiology, July 2005, p. 3129-3135, Vol. 43, No. 7
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.7.3129-3135.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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