Korean J Lab Med.  2009 Oct;29(5):448-454. 10.3343/kjlm.2009.29.5.448.

Comparison of 3 Phenotypic-detection Methods for Identifying Plasmid-mediated AmpC beta-lactamase-producing Escherichia coli, Klebsiella pneumoniae, and Proteus mirabilis Strains

• Lee W
• Jung B
• Hong SG
• Song W
• Jeong SH
• Lee K
• Kwak HS

• KMID: 1077421
• DOI: http://doi.org/10.3343/kjlm.2009.29.5.448

Abstract

BACKGROUND
Plasmid-mediated AmpC beta-lactamases (PABLs) have been detected in the strains of Escherichia coli, Klebsiella spp., Proteus mirabilis, and Salmonella spp. PABLs may be difficult to detect and might interfere in the therapeutic and infection-control processes. Although several PABL-detection methods based on phenotypes have been reported, the Clinical and Laboratory Standards Institute currently does not recommend a routine detection method for PABLs. The aim of this study is to compare the performances of 3 phenotypic PABL detection methods.
METHODS
Total 276 non-duplicated clinical isolates of E. coli (N=97), K. pneumoniae (N=136), and P. mirabilis (N=43) were collected from 14 hospitals in Korea between April and June 2007 in a non-consecutive and non-random manner. Multiplex PCR was performed to detect the PABL genes. Further, 3 phenotypic detection methods-cephamycin-Hodge test, Tris-EDTA (TE) disk test, and combination-disk test with 3-aminophenylboronic acid (BA)-were performed using cefoxitin and cefotetan disks. RESULTS: PABL genes were detected by multiplex PCR in 122/276 isolates, including 14/97 E. coli, 105/136 K. pneumoniae, and 3/43 P. mirabilis isolates. The combination-disk test with BA showed higher sensitivity (98.4%), specificity (92.2%), and efficiency (96.3%) than the cephamycin-Hodge (76.2%, 96.1%, and 88.6%, respectively) and the TE-disk (80.3%, 91.6%, and 87.9%, respectively) tests.
CONCLUSIONS
The combination-disk test with BA is a simple, efficient, and interpretable test that can be applicable in clinical laboratories involved in the detection of PABLs in clinical isolates of E. coli, K. pneumoniae, and P. mirabilis.

Keyword

AmpC beta-lactamase; Boronic acid; Hodge test; Tris-EDTA disk test

MeSH Terms

Anti-Bacterial Agents/pharmacology
Bacterial Proteins/*analysis
Cefotetan/pharmacology
Cefoxitin/pharmacology
Disk Diffusion Antimicrobial Tests/*methods
Escherichia coli/genetics/*isolation & purification
Humans
Klebsiella pneumoniae/genetics/*isolation & purification
Phenotype
Plasmids
Proteus mirabilis/genetics/*isolation & purification
Sensitivity and Specificity
beta-Lactamases/*analysis

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