ORIGINAL ARTICLES

Detection of plasmid-mediated AmpC in Escherichia coli, Klebsiella pneumoniae and Proteus mirabilis

T Y Tan¹, S Y Ng¹, L Teo², Y Koh² and C H Teok²

¹ Division of Laboratory Medicine, Changi General Hospital, Singapore
² School of Chemical and Life Sciences, Singapore Polytechnic, Singapore

Correspondence to:
Thean Yen Tan, Division of Laboratory Medicine, Changi General Hospital, 2 Simei Street 3, Singapore 529889; thean_yen_tan{at}cgh.com.sg

Aims: This study investigated the prevalence of plasmid-mediated AmpC production in selected clinical isolates of Escherichia coli, Klebsiella species and Proteus mirabilis, and compared the results of boronic acid disc screening with conventional susceptibility testing for the detection of AmpC-positive isolates.

Methods: E coli, Klebsiella species and P mirabilis with reduced susceptibility to amoxycillin-clavulanate, cefuroxime and cephalexin, but without phenotypic evidence of extended-spectrum β-lactamases were screened for AmpC activity using enzyme-extraction methods. The presence of plasmid-mediated ampC was determined by multiplex PCR. Antibiotic susceptibilities were determined using both disc and dilution-based methods. A disc-based screening method for detection of AmpC-producing strains was evaluated using boronic acid as an inhibitor of AmpC, and cefoxitin as the antibiotic substrate.

Results: Plasmid-mediated ampC was present in 26% of study isolates, with CMY-like enzymes detected predominantly in E coli and DHA-like enzymes predominantly in Klebsiella pneumoniae. Current susceptibility methods failed to detect a significant proportion of plasmid-mediated AmpC-producing isolates, with 33% of such strains interpreted as susceptible to third-generation cephalosporins using current Clinical Laboratory Standards Institute breakpoints. The boronic acid disc method showed sensitivity and specificity of 90% and 98% respectively in detecting AmpC-positive isolates.

Conclusion: The prevalence of plasmid-mediated ampC was high in the study population, and may be missed by conventional susceptibility testing methods. Inhibitor-based screening methods would improve detection of this emerging resistance phenotype.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

• Tsakris, A., Kristo, I., Poulou, A., Themeli-Digalaki, K., Ikonomidis, A., Petropoulou, D., Pournaras, S., Sofianou, D. (2009). Evaluation of Boronic Acid Disk Tests for Differentiating KPC-Possessing Klebsiella pneumoniae Isolates in the Clinical Laboratory. J. Clin. Microbiol. 47: 362-367 [Abstract] [Full Text]  
• Jacoby, G. A. (2009). AmpC {beta}-Lactamases. Clin. Microbiol. Rev. 22: 161-182 [Abstract] [Full Text]  
• Tan, T. Y., Ng, L. S. Y., He, J., Koh, T. H., Hsu, L. Y. (2009). Evaluation of Screening Methods To Detect Plasmid-Mediated AmpC in Escherichia coli, Klebsiella pneumoniae, and Proteus mirabilis. Antimicrob. Agents Chemother. 53: 146-149 [Abstract] [Full Text]