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Journal of Clinical Microbiology, November 2009, p. 3409-3412, Vol. 47, No. 11
0095-1137/09/$08.00+0     doi:10.1128/JCM.01141-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Broth Microdilution Method To Detect Extended-Spectrum β-Lactamases and AmpC β-Lactamases in Enterobacteriaceae Isolates by Use of Clavulanic Acid and Boronic Acid as Inhibitors {triangledown}

Seok Hoon Jeong,1 Wonkeun Song,2* Jae-Seok Kim,2 Han-Sung Kim,2 and Kyu Man Lee2

Department of Laboratory Medicine and Research Institute of Bacterial Resistance, Yonsei University College of Medicine,1 Department of Laboratory Medicine, Hallym University College of Medicine, Seoul, Republic of Korea2

Received 11 June 2009/ Returned for modification 6 August 2009/ Accepted 14 August 2009

This study was designed to evaluate the performance of the broth microdilution (BMD) method to detect production of extended-spectrum β-lactamases (ESBLs) and AmpC β-lactamases in Enterobacteriaceae by using clavulanic acid (CA) and boronic acid (BA) as ESBL and AmpC β-lactamase inhibitors, respectively. A total of 100 clinical isolates of Enterobacteriaceae were analyzed. Mueller-Hinton broth containing serial twofold dilutions of cefotaxime (CTX), ceftazidime (CAZ), aztreonam (ATM), or cefepime (FEP) with or without either or both CA and BA was prepared. An eightfold or greater decrease in the MIC of CTX, CAZ, ATM, or FEP in the presence of CA and BA was considered a positive result for ESBL and plasmid-mediated AmpC β-lactamase (PABL), respectively. In tests with CA, expanded-spectrum β-lactams containing BA (CTX-BA, CAZ-BA, ATM-BA, and FEP-BA) showed higher positive rates in detecting ESBL producers than those without BA. The combination of CTX- and CAZ-based BMD tests with CA and BA showed sensitivity and specificity of 100% for the detection of ESBLs and PABLs. The BMD testing could be applicable for routine use in commercially available semiautomated systems for the detection of ESBLs and PABLs in Enterobacteriaceae.

* Corresponding author. Mailing address: Department of Laboratory Medicine, Hallym University College of Medicine, 948-1 Daerim 1-Dong, Youngdeungpo-Gu, Seoul 150-950, Republic of Korea. Phone: 82-2-829-5259. Fax: 82-2-847-2403. E-mail: swonkeun{at}hallym.or.kr

{triangledown} Published ahead of print on 26 August 2009.

Journal of Clinical Microbiology, November 2009, p. 3409-3412, Vol. 47, No. 11
0095-1137/09/$08.00+0     doi:10.1128/JCM.01141-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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