Simple Disk-Based Method for Detection of
            <i>Klebsiella pneumoniae</i>
            Carbapenemase-Type β-Lactamase by Use of a Boronic Acid Compound

Doi, Yohei; Potoski, Brian A.; Adams-Haduch, Jennifer; Sidjabat, Hanna E.; Pasculle, A. William; Paterson, David L.

doi:10.1128/jcm.01408-08

Cited by 133 publications

(93 citation statements)

References 23 publications

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“…Another culture-based method, used for detection of carbapenemases among A. baumannii is represented by KPC combined disk assay. This technique utilizes boronic acid compound (BAC) as KPC inhibitor [96,97].…”

Section: Phenotypic Methodsmentioning

confidence: 99%

Acinetobacter baumannii: biology and drug resistance — role of carbapenemases

Nowak

Paluchowska

2015

Folia Histochem Cytobiol.

107

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Acinetobacter baumannii is a Gram-negative, glucose-non-fermenting, oxidase-negative coccobacillus, most commonly associated with the hospital settings. The ability to survive in adverse environmental conditions as well as high level of natural and acquired antimicrobial resistance make A. baumannii one of the most important nosocomial pathogens. While carbapenems have long been considered as antimicrobials of last-resort, the rates of clinical A. baumannii strains resistant to these antibiotics are increasing worldwide. Carbapenem resistance among A. baumannii is conferred by coexisting mechanisms including: decrease in permeability of the outer membrane, efflux pumps, production of beta-lactamases, and modification of penicillin-binding proteins. The most prevalent mechanism of carbapenem resistance among A. baumannii is associated with carbapenem-hydrolysing enzymes that belong to Ambler class D and B beta-lactamases. In addition, there have also been reports of resistance mediated by selected Ambler class A carbapenemases among A. baumannii strains. Resistance determinants in A. baumannii are located on chromosome and plasmids, while acquisition of new mechanisms can be mediated by insertion sequences, integrons, transposons, and plasmids. Clinical relevance of carbapenem resistance among strains isolated from infected patients, carriers and hospital environment underlines the need for carbapenemase screening. Currently available methods vary in principle, accuracy and efficiency. The techniques that deserve particular attention belong to both easily accessible unsophisticated methods as well as advanced techniques based on mass spectrometry or molecular biology. While carbapenemases limit the therapeutic options in A. baumannii infections, studies concerning novel beta-lactamase inhibitors offer a new insight into effective therapy.

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Section: Phenotypic Methodsmentioning

confidence: 99%

Acinetobacter baumannii: biology and drug resistance — role of carbapenemases

Nowak

Paluchowska

2015

Folia Histochem Cytobiol.

107

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“…Unexpectedly, we discovered unusual inhibition of organisms possessing KPC by 3-aminophenylboronic acid (APB), which may be attributed solely to the presence of this carbapenemase (25). Afterwards, Doi et al and, in a subsequent study with a larger series of strains, Tsakris et al demonstrated the overwhelming effectiveness of APB for the detection of Klebsiella pneumoniae possessing KPC (7,32). Given these promising results, we decided to explore the potential of APB methods as confirmatory tests for class A carbapenemase detection.…”

mentioning

confidence: 94%

Sensitive Screening Tests for Suspected Class A Carbapenemase Production in Species of Enterobacteriaceae

et al. 2009

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The detection of class A serine-carbapenemases among species of Enterobacteriaceae remains a challenging issue. Methods of identification for routine use in clinical microbiology laboratories have not been standardized to date. We developed a novel screening methodology suitable for countries with high basal levels of carbapenem resistance due to non-carbapenemase-mediated mechanisms and standardized several simple confirmatory methods that allow the recognition of bacteria producing class A carbapenemases, including KPC, Sme, IMI, NMC-A, and GES, by using boronic acid (BA) derivatives. A total of 28 genetically unrelated Enterobacteriaceae strains producing several class A carbapenemases were tested. Thirty-eight genetically unrelated negative controls were included. The isolates were tested against imipenem (IPM), meropenem (MEM), and ertapenem (ETP) by MIC and disk diffusion assays in order to select appropriate tools to screen for suspected carbapenemase production. It was possible to differentiate class A carbapenemase-producing bacteria from non-carbapenemase-producing bacteria by using solely the routine IPM susceptibility tests. The modified Hodge test was evaluated and found to be highly sensitive, although false-positive results were documented. Novel BA-based methods (a double-disk synergy test and combined-disk and MIC tests) using IPM, MEM, and ETP, in combination with 3-aminophenylboronic acid as an inhibitor, were designed as confirmatory tools. On the basis of the performance of these methods, a sensitive flow chart for suspicion and confirmation of class A carbapenemase production in species of Enterobacteriaceae was designed. By using this methodology, isolates producing KPC, GES, Sme, IMI, and NMC-A carbapenemases were successfully distinguished from those producing other classes of ␤-lactamases (extended-spectrum ␤-lactamases, AmpCs, and metallo-␤-lactamases, etc). These methods will rapidly provide useful information needed for targeting antimicrobial therapy and appropriate infection control.

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“…(Table 3). However, recently, a study showed that 3-aminophenylboronic acid inhibited the activity of KPC-type β -lactamase [24]. Doi et al [24] reported that 3-aminophenylboronic acid could be used for detecting the expression of KPC-type β -lactamase in K.…”

Section: Resultsmentioning

confidence: 99%

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

et al. 2009

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Background : Plasmid-mediated AmpC β -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. (Korean J Lab Med 2009;29:448-54)

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Simple Disk-Based Method for Detection of Klebsiella pneumoniae Carbapenemase-Type β-Lactamase by Use of a Boronic Acid Compound

Cited by 133 publications

References 23 publications

Acinetobacter baumannii: biology and drug resistance — role of carbapenemases

Acinetobacter baumannii: biology and drug resistance — role of carbapenemases

Sensitive Screening Tests for Suspected Class A Carbapenemase Production in Species of Enterobacteriaceae

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

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