Detection of Extended-Spectrum β-Lactamases by Using Boronic Acid as an AmpC β-Lactamase Inhibitor in Clinical Isolates of
            <i>Klebsiella</i>
            spp. and
            <i>Escherichia coli</i>

Song, Wonkeun; Bae, Il Kwon; Lee, You-Nae; Lee, Chae-Hoon; Lee, Sangeun; Jeong, Seok Hoon

doi:10.1128/jcm.02322-06

Cited by 72 publications

(57 citation statements)

References 7 publications

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“…Addition of 3-aminophenyl boronic acid on antibiotic-containing disks (cefotaxime/ceftazidime with or without clavulanate) has been reported to potentiate the sensitivity and specificity of the combination disk method when this method is applied to organisms with chromosomal [74] or plasmid-mediated [75] AmpC production.…”

Section: Detection Methods With Boronic Acidmentioning

confidence: 99%

Extended-spectrumβ-Lactamases: Implications for the Clinical Laboratory and Therapy

2008

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Section: Detection Methods With Boronic Acidmentioning

confidence: 99%

Extended-spectrumβ-Lactamases: Implications for the Clinical Laboratory and Therapy

2008

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“…Hodge and imipenem and EDTA-sodium mercaptoacetic acid double-disc synergy (IEDDS) tests were performed on agar plates as described previously (Lee et al, 2001(Lee et al, , 2010a. The MICs of imipenem and meropenem were determined using Etest strips (AB Biodisk) on Mueller-Hinton agar plates with or without a fixed concentration of clavulanic acid (CA; 4 mg ml 21 ; Sigma-Aldrich) as an inhibitor of ESBLs, aminophenylboronic acid (APB; 300 mg ml 21 ; Sigma-Aldrich) as an inhibitor of AmpC b-lactamase, or Phe-Arg-b-naphthylamide (PAbN; 20 mg ml 21 ; SigmaAldrich) as an efflux pump inhibitor (Song et al, 2007).…”

Section: Methodsmentioning

confidence: 99%

Resistance to carbapenems in sequence type 11 Klebsiella pneumoniae is related to DHA-1 and loss of OmpK35 and/or OmpK36

Shin

Bae

Kim

et al. 2012

Journal of Medical Microbiology

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Resistance to carbapenems in sequence type 11 Klebsiella pneumoniae is related to DHA-1 and loss of OmpK35 and/or OmpK36 This study investigated the molecular mechanisms of carbapenem resistance in clinical isolates of Klebsiella pneumoniae from Korea and the clinical outcomes of resistant K. pneumoniae infection. Sixteen K. pneumoniae isolates showing resistance to carbapenems collected from a tertiary-care hospital were examined for the mechanisms of carbapenem resistance. PCR and sequencing experiments detected the bla DHA-1 AmpC b-lactamase gene in all 16 clinical isolates, whilst the bla genes of extended-spectrum b-lactamases were detected in 12 of 16 isolates. SDS-PAGE experiments indicated that all the isolates lacked the 35 kDa and/or 36 kDa outer-membrane proteins (OMPs). Sequence analysis of the corresponding OMP genes revealed various alterations. PFGE analysis demonstrated that there were no major clonal relationships among the K. pneumoniae isolates. However, multilocus sequence typing experiments showed that all isolates shared the same sequence type (ST), ST11, except for one isolate of ST48. The crude mortality rate of infected patients was 81.8 %. Carbapenem resistance was mainly due to a combination of DHA-1 AmpC b-lactamase coupled with the loss of OmpK35 and/or OmpK36 and was associated with poor clinical outcome. INTRODUCTIONCarbapenems are widely used and are often the only therapeutic option for treating serious infections caused by Klebsiella pneumoniae that produces extended-spectrum b-lactamases (ESBLs) and/or AmpC b-lactamases. The recent emergence of carbapenem-resistant K. pneumoniae in various countries is therefore worrying, as antimicrobial treatment options are limited.Carbapenem resistance in K. pneumoniae has been ascribed mainly to enzymic degradation by plasmid-borne carbapenemases of classes A (GES-2, -4, -5 and -6, and KPC enzymes), B (IMP-1 and -8, and VIM-1, -2 and -12) and D (OXA-48) (Poirel et al., 2004a;Queenan & Bush, 2007). However, some K. pneumoniae strains have acquired carbapenem resistance despite their lack of carbapenemase production. Loss of outer-membrane proteins (OMPs), including OmpK35 and OmpK36, plays a role in carbapenem resistance in K. pneumoniae strains harbouring ESBLs METHODSPatients and strains. This study was conducted between 2007 and 2008 at a tertiary-care teaching hospital with 2000 beds in Seoul, Korea. During this period, K. pneumoniae was isolated from 3278 patients, 16 (0.5 %) of which had K. pneumoniae clinical isolates that exhibited resistance to carbapenems. The isolates were identified using a Vitek GNI card (bioMérieux). Clinical characteristics were investigated through electronic medical record review. Data including age, sex, underlying disease, number of days from hospital admission to identification of K. pneumoniae infection, prior antibiotic treatment, anatomical site of isolation and clinical outcome were obtained.Antimicrobial susceptibility testing. Antibiotic-containing discs (Becton Dickinson) were used for routine an...

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“…The MBL Etest (AB Biodisk, Solna, Sweden) and the combined-disk test with imipenem and EDTA (10) were used to screen for the production of class B carbapenemases. ESBL production was tested using the CLSI confirmatory test and a modification, which uses clavulanate in combination with boronic acid (26). Isolates were phenotypically tested for the presence of AmpC ␤-lactamase by determining cefoxitin and imipenem MICs in agar with and without 200 g/ml cloxacillin (23) and by using the AmpC detection Etest strips (AB Biodisk), which contain cefotetan with or without cloxacillin.…”

Section: Methodsmentioning

confidence: 99%

Evaluation of Boronic Acid Disk Tests for Differentiating KPC-Possessing Klebsiella pneumoniae Isolates in the Clinical Laboratory

et al. 2009

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The worldwide increase in the occurrence and dissemination of KPC ␤-lactamases among gram-negative pathogens makes critical the early detection of these enzymes. Boronic acid disk tests using different antibiotic substrates were evaluated for detection of KPC-possessing Klebsiella pneumoniae isolates. A total of 57 genotypically confirmed KPC-possessing K. pneumoniae isolates with varying carbapenem MICs were examined. To measure the specificity of the tests, 106 non-KPC-possessing isolates (89 K. pneumoniae and 17 Escherichia coli isolates) were randomly selected among those exhibiting reduced susceptibility to cefoxitin, expanded-spectrum cephalosporins, or carbapenems. As many as 56, 53, and 40 of the non-KPC-possessing isolates harbored extended-spectrum ␤-lactamases, metallo-␤-lactamases, and plasmidmediated AmpC ␤-lactamases, respectively. By use of CLSI methodology and disks containing imipenem, meropenem, or cefepime, either alone or in combination with 400 g of boronic acid, all 57 KPC producers gave positive results (sensitivity, 100%) whereas all 106 non-KPC producers were negative (specificity, 100%). The meropenem duplicate disk with or without boronic acid demonstrated the largest differences in inhibition zone diameters between KPC producers and non-KPC producers. By use of disks containing ertapenem, all isolates were correctly differentiated except for five AmpC producers that gave falsepositive results (sensitivity, 100%; specificity, 95.3%). These practical and simple boronic acid disk tests promise to be very helpful for the accurate differentiation of KPC-possessing K. pneumoniae isolates, even in regions where different broad-spectrum ␤-lactamases are widespread.

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Detection of Extended-Spectrum β-Lactamases by Using Boronic Acid as an AmpC β-Lactamase Inhibitor in Clinical Isolates of Klebsiella spp. and Escherichia coli

Cited by 72 publications

References 7 publications

Extended-spectrumβ-Lactamases: Implications for the Clinical Laboratory and Therapy

Extended-spectrumβ-Lactamases: Implications for the Clinical Laboratory and Therapy

Resistance to carbapenems in sequence type 11 Klebsiella pneumoniae is related to DHA-1 and loss of OmpK35 and/or OmpK36

Evaluation of Boronic Acid Disk Tests for Differentiating KPC-Possessing Klebsiella pneumoniae Isolates in the Clinical Laboratory

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