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Journal of Clinical Microbiology, February 2009, p. 362-367, Vol. 47, No. 2
0095-1137/09/$08.00+0     doi:10.1128/JCM.01922-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Evaluation of Boronic Acid Disk Tests for Differentiating KPC-Possessing Klebsiella pneumoniae Isolates in the Clinical Laboratory{triangledown}

Athanassios Tsakris,1* Ioulia Kristo,2 Aggeliki Poulou,3 Katerina Themeli-Digalaki,4 Alexandros Ikonomidis,2 Dimitra Petropoulou,5 Spyros Pournaras,2 and Danai Sofianou6

Department of Microbiology, Medical School, University of Athens, Athens,1 Department of Microbiology, Medical School, University of Thessaly, Larissa,2 Department of Microbiology, Serres General Hospital, Serres,3 Department of Microbiology, Tzaneion General Hospital, Piraeus,4 Department of Microbiology, Saint Panteleimon General Hospital, Nicea,5 Department of Microbiology, Hippokration University Hospital, Thessaloniki, Greece6

Received 5 October 2008/ Returned for modification 17 November 2008/ Accepted 4 December 2008

The worldwide increase in the occurrence and dissemination of KPC β-lactamases among gram-negative pathogens makes critical the early detection of these enzymes. Boronic acid disk tests using different antibiotic substrates were evaluated for detection of KPC-possessing Klebsiella pneumoniae isolates. A total of 57 genotypically confirmed KPC-possessing K. pneumoniae isolates with varying carbapenem MICs were examined. To measure the specificity of the tests, 106 non-KPC-possessing isolates (89 K. pneumoniae and 17 Escherichia coli isolates) were randomly selected among those exhibiting reduced susceptibility to cefoxitin, expanded-spectrum cephalosporins, or carbapenems. As many as 56, 53, and 40 of the non-KPC-possessing isolates harbored extended-spectrum β-lactamases, metallo-β-lactamases, and plasmid-mediated AmpC β-lactamases, respectively. By use of CLSI methodology and disks containing imipenem, meropenem, or cefepime, either alone or in combination with 400 µg of boronic acid, all 57 KPC producers gave positive results (sensitivity, 100%) whereas all 106 non-KPC producers were negative (specificity, 100%). The meropenem duplicate disk with or without boronic acid demonstrated the largest differences in inhibition zone diameters between KPC producers and non-KPC producers. By use of disks containing ertapenem, all isolates were correctly differentiated except for five AmpC producers that gave false-positive results (sensitivity, 100%; specificity, 95.3%). These practical and simple boronic acid disk tests promise to be very helpful for the accurate differentiation of KPC-possessing K. pneumoniae isolates, even in regions where different broad-spectrum β-lactamases are widespread.

* Corresponding author. Mailing address: Department of Microbiology, Medical School, University of Athens, 11527 Athens, Greece. Phone: 30 210 7462011. Fax: 30 210 7462210. E-mail: atsakris{at}med.uoa.gr

{triangledown} Published ahead of print on 10 December 2008.

Journal of Clinical Microbiology, February 2009, p. 362-367, Vol. 47, No. 2
0095-1137/09/$08.00+0     doi:10.1128/JCM.01922-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.



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