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Abstract
Background
Accurate detection of organisms producing extended-spectrum β-lactamase (ESBL) and AmpC β-lactamase is very important for treatment of patients. However, unlike the ESBL confirmatory test, there are no guidelines for detection of organisms producing AmpC β-lactamase. We evaluated a detection method using boronic acid (BA) for ESBL and AmpC β-lactamase.
Methods
Clinical isolates of Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, and Proteus mirabilis showing intermediate resistance or resistance to cefoxitin (FOX) or positive for ESBL were tested. A ≥5 mm increase in zone diameter of ceftazidime/clavulanic acid/BA (CAZ/CA/BA) and/or cefotaxime/clavulanic acid/BA (CTX/CA/BA) versus CAZ/BA and/or CTX /BA was considered positive for ESBL. Likewise, a ≥5 mm increase in zone diameter of FOX/BA and/or cefotetan/BA (CTT/BA) versus FOX and/or CTT alone was considered positive for AmpC β-lactamase.
Results
Among 622 clinical isolates, ESBL positive rates by the CLSI ESBL confirmatory test or by the BA method were 18.1% or 18.4% for E. coli, 38.3% or 40.4% for K. pneumoniae, 8.7% or 8.7% for K. oxytoca, and 14.8% or 14.8% for P. mirabilis, respectively. AmpC β-lactamase positive rates using the BA method were 3.7% for E. coli, 33.3% for K. pneumoniae, 0% for K. oxytoca, and 7.4% for P. mirabilis. The detection rates of coproducing ESBL and AmpC β-lactamase were 2.4% in E. coli 27.1% in K. pneumoniae, and 3.7% in P. mirabilis.
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Fig. 1.
Representative results using the Clinical and Laboratory Standards Institute extended-spectrum β-lactamase (ESBL) confirmatory test and AmpC disk test without and with boronic acid (two disks positioned in right side on media). Abbreviations: FOX, cefoxitin; CAZ, ceftazidime; CV, clavulanic acid; CTX, cefotaxime; CTX, cefotaxime. Left media shows AmpC positive, center media shows ESBL positive, and right media shows that isolate is negative in CLSI method and positive in boronic acid disk.
Table 1.
Comparison of CLSI ESBL confirmatory tests in Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, and Proteus mirabilis
Table 2.
Comparative results in E. coli, K. pneumoniae, K. oxytoca and P. mirabilis by combination with the boronic acid
Table 3.
Prevalence of AmpC and ESBL in E. coli, K. pneumoniae, K. oxytoca and P. mirabilis by combination with the boronic acid
Table 4.
Antimicrobial susceptibilities of E. coli, K. pneumoniae, K. oxytoca, and P. mirabilis according to the presence of AmpC and/or ESBL
Abbreviations: ESBL, extended-spectrum β-lactamase; SXT, cotrimoxazole; CIP, ciprofloxacin; AMK, amikacin; GM, gentamicin; TOB, tobramycin; MAN, cefamandole; CFS, cefoperazone/sulbactam; PPT, piperacillin/tazobactam; ATM, aztreonam; CTX, cefotaxime; CAZ, ceftazidime; FEP, cefepime; FOX, cefoxitin; CTT, cefotetan; IPM, imipenem.
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