Evaluation of the modified carbapenem inactivation method and sodium mercaptoacetate-combination method for the detection of metallo-β-lactamase production by carbapenemase-producing Enterobacteriaceae

Yamada, Kageto; Kashiwa, Machiko; Arai, Katsumi; Nagano, Noriyuki; Saito, Ryoichi

doi:10.1016/j.mimet.2016.11.013

Cited by 27 publications

(26 citation statements)

References 9 publications

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“…The CIM has been evaluated worldwide by methods similar to those used in our study (21)(22)(23). Some papers have pointed out that the CIM cannot detect some portion of carbapenemase producers or bacterial species, and several modified CIMs have been reported (24,25). In our study, however, the CIM could effectively detect CPE strains with high specificity, even strains with low carbapenem MICs, such as IMP producers.…”

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confidence: 79%

Suitability of Carbapenem Inactivation Method (CIM) for Detection of IMP Metallo-β-Lactamase-Producing Enterobacteriaceae

et al. 2017

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Carbapenem-resistant Enterobacteriaceae (CRE) strains have become globally distributed in the past decade, resulting in concern over the control of hospital infections and antimicrobial therapies (1, 2). The majority of CRE isolates are carbapenemase-producing Enterobacteriaceae (CPE) strains, so early detection of CPE strains is essential for providing optimal antimicrobial therapies and preventing horizontal transmission. In 2015, van der Zwaluw et al. reported the carbapenem inactivation method (CIM), a new method for detecting carbapenemase producers with high sensitivity and specificity (3). However, it was unclear whether CIM can identify IMP producers with a low carbapenem MIC or non-CPE strains that are highly carbapenem resistant. In this study, we evaluated whether CIM can identify CPE strains independently of their carbapenem MICs.Were used 233 CPE and 51 non-CPE strains isolated in general hospitals across Japan from 2012 to 2016 and stocked in our laboratory. The strains were collected from blood, sputum, wounds, urine, and feces. Their antibiotic susceptibilities were determined by the agar dilution method in accordance with the recommendations of the CLSI (4). The CPE strains included 191 IMP, 20 KPC, 17 NDM, and 5 OXA-48-like (OXA-48 and OXA-244) producers. The non-CPE strains included 31 extended-spectrum beta-lactamase (CTX-M, SHV, and TEM) producers and 5 plasmid-mediated AmpC ␤-lactamase (DHA, CMY, and CFE) producers. They were identified by DNA sequence amplification as previously described (5-10). Of the non-CPE strains, 13 highly carbapenem-resistant strains were identified as producing cephalosporinases and lacking porin function by SDS-PAGE, DNA sequencing, and quantitative reverse transcription-PCR (11).The CIM was conducted as previously described (3). The isolates were cultured on Mueller-Hinton agar (MHA) plates. A full 10-l inoculation loop of each strain was suspended in 400 l of sterile distilled water, and a 10-g meropenem (MEM) susceptibilitytesting disk (E-DF85; EIKEN) was immersed in the solution. After incubation at 35°C for 2 h, the disk was removed and placed on an MHA plate inoculated with a 0.5 McFarland standard of Escherichia coli strain ATCC 25922 with a sterile cotton swab. Finally, the plate was incubated overnight at 35°C and the inhibition zone around each disk was measured. Inhibition circles Ͻ10 mm in diameter were judged to indicate CIM positivity.The CIM showed a sensitivity of 100% (233/233) for CPE strains and a specificity of 96.1% (49/51) for non-CPE strains (Table 1). The MICs of MEM for the 191 IMP producers ranged from 0.125 to 32 g/ml (MIC 50 ϭ 0.5 g/ml, MIC 90 ϭ 2 g/ml), and those of imipenem (IPM) ranged from 0.06 to 8 g/ml (MIC 50 ϭ 0.125 g/ml, MIC 90 ϭ 0.5

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confidence: 79%

Suitability of Carbapenem Inactivation Method (CIM) for Detection of IMP Metallo-β-Lactamase-Producing Enterobacteriaceae

et al. 2017

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“…While we observed two false-positive results with OXA-48producing isolates (specifically, OXA-232 enzymes) when EDTA was added at a final concentration of 5 mM, the zones of growth inhibition in the presence of EDTA for both isolates were less than those observed for the MBL-producing isolates and, importantly, both isolates were mCIM positive and eCIM negative upon repeat testing. Recently, two phenotypic methods for differentiating carbapenemase classes have been described: SMA-mCIM and CIMplus (13,15). SMA-mCIM uses an alternative MBL inhibitor, sodium mercaptoacetate, in the same format as the mCIM and demonstrated a sensitivity and specificity of 100% (13).…”

Section: Discussionmentioning

confidence: 99%

“…Recently, two phenotypic methods for differentiating carbapenemase classes have been described: SMA-mCIM and CIMplus (13,15). SMA-mCIM uses an alternative MBL inhibitor, sodium mercaptoacetate, in the same format as the mCIM and demonstrated a sensitivity and specificity of 100% (13). CIMplus employs two specific carbapenemase inhibitors (phenylboronic acid for class A enzymes and EDTA for class B enzymes) and a decision tree to differentiate between class A, B, and D enzymes (15).…”

Section: Discussionmentioning

confidence: 99%

“…Recently, a modified variant of the CIM, mCIM, was developed for phenotypic detection of CP-CRE isolated in culture (11). The mCIM is highly sensitive and specific (11,13); however, it does not differentiate carbapenemase-producing Enterobacteriaceae expressing serine carbapenemases (i.e., class A and D enzymes) from those elaborating MBLs. This present study describes the development and evaluation of the EDTA-mCIM, eCIM, which permits differentiation of serine enzymes and MBLs in a format compatible with the mCIM.…”

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confidence: 99%

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EDTA-Modified Carbapenem Inactivation Method: a Phenotypic Method for Detecting Metallo-β-Lactamase-Producing Enterobacteriaceae

et al. 2019

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The increase in the prevalence and impact of infections caused by carbapenemase-producing Enterobacteriaceae is a global health concern. Therefore, rapid and accurate methods to detect these organisms in any clinical microbiology laboratory, including those in resource-limited settings, are essential to prevent and contain their spread. It is also important to differentiate between serine-and metaldependent carbapenemases elaborated by carbapenemase-producing isolates for epidemiologic, infection control and prevention, and therapeutic purposes. Here, we describe the development and evaluation of the EDTA-modified carbapenem inactivation method (eCIM), an assay for discriminating between serine-and metaldependent (i.e., metallo-␤-lactamases [MBLs]) carbapenemases when used in conjunction with the modified carbapenem inactivation method (mCIM). The eCIM had an overall sensitivity and specificity of 100% and was adopted by the Clinical and Laboratory Standards Institute as a method to use in combination with the mCIM to identify MBL-producing Enterobacteriaceae.

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“…Carbapenems are considered the most reliably active treatment option against severe infections caused by multidrug-resistant and extensively drug-resistant Gram-negative bacilli (2)(3)(4). In Enterobacteriaceae, carbapenem resistance develops via two main mechanisms: (i) acquisition of carbapenemase genes that encode enzymes capable of degrading carbapenems, or (ii) overexpression of β-lactamases that possess very weak affinity for carbapenems coupled with modifications in permeability of porins and/or expression of efflux pumps (1).…”

Section: Introductionmentioning

confidence: 99%

Phenotypic and molecular identification of carbapenemase-producing Enterobacteriaceae - challenges in diagnosis and treatment

Főldes¹,

Bilca

Székely

2018

Revista Romana De Medicina De Laborator

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Introduction: Infections due to carbapenemase-producing carbapenem-resistant Enterobacteriaceae (CPCRE) are an emerging global public health threat. The purpose of this study was to investigate phenotypic and genotypic features of CP-CRE strains isolated from hospitalized patients. Material and methods: Between 1st of January - 1st of July 2017, in the Department of Microbiology, “Dr. Constantin Opriş” County Emergency Hospital Baia Mare, Romania, 1110 strains of Enterobacteriaceae were isolated from bronchial secretions, urine, wounds and blood cultures. Bacterial identification and antimicrobial susceptibility tests were performed by conventional methods, Vitek 2 Compact and M.I.C.E. strips. We analysed all Enterobacteriaceae strains non-susceptible to carbapenems according to CLSI 2017 criteria. The modified Hodge test (MHT), the modified carbapenem inactivation method (mCIM) and the combination disks test (KPC, MBL, OXA-48 Confirm kit, Rosco Diagnostica) were used for phenotypic confirmation, whereas a multiplex PCR assay for genes blaKPC, blaNDM and blaOXA-48 was used for genetic confirmation. Results: 19 non-duplicate strains isolated from 16 patients were phenotypically identified as CP-CRE: Klebsiella pneumoniae (n=14), Escherichia coli (n=2), Providencia stuartii (n=2) and Serratia marcescens (n=1). Most strains were isolated from bronchial secretions (n=9). The carbapenem-hydrolizing enzymes were identified by the combination disks test as: KPC (n=9), OXA-48-like (n=5) and MBL (n=5). Molecular confirmation was performed in 18 phenotypically positive isolates with 100% concordant results with mCIM and combination disks test. Discrepant results were noticed with the MHT in case of 4 NDM-producers confirmed by PCR. All CP-CRE strains were resistant to all tested cephems. Three out of 9 K. pneumoniae strains tested against colistin were found resistant. Conclusions: The most common carbapenemase detected was KPC. Therapeutic options were limited in all positive cases. Rapid and reliable detection of CP-CRE is critical for preventing the spread of these pathogens

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Evaluation of the modified carbapenem inactivation method and sodium mercaptoacetate-combination method for the detection of metallo-β-lactamase production by carbapenemase-producing Enterobacteriaceae

Cited by 27 publications

References 9 publications

Suitability of Carbapenem Inactivation Method (CIM) for Detection of IMP Metallo-β-Lactamase-Producing Enterobacteriaceae

Suitability of Carbapenem Inactivation Method (CIM) for Detection of IMP Metallo-β-Lactamase-Producing Enterobacteriaceae

EDTA-Modified Carbapenem Inactivation Method: a Phenotypic Method for Detecting Metallo-β-Lactamase-Producing Enterobacteriaceae

Phenotypic and molecular identification of carbapenemase-producing Enterobacteriaceae - challenges in diagnosis and treatment

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