Retrospective and prospective evaluation of the Carbapenem inactivation method for the detection of carbapenemase-producing Enterobacteriaceae (original) (raw)
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The Journal of antimicrobial chemotherapy, 2018
Fast and accurate diagnostic tests to identify carbapenemase-producing Enterobacteriaceae (CPE) are mandatory for proper antimicrobial therapy and implementing infection control measures. Here, we have developed a rapid Carbapenem Inactivation Method (rCIM) for CPE detection. The rCIM consists of the incubation of a potential carbapenemase producer with meropenem discs and use of the resulting supernatant to challenge a susceptible indicator strain. Growth of the indicator strain is monitored using a nephelometer. The performances of the rCIM were compared with the CIM and Carba NP tests using a collection of 113 well-characterized carbapenem-resistant enterobacterial isolates, including 85 carbapenemase producers and 28 non-carbapenemase producers. In addition, rCIM was compared with the Carba NP test and PCR sequencing in a prospective analysis of 101 carbapenem-resistant enterobacterial isolates addressed to the French National Reference Center for Antimicrobial Resistance in Jul...
PLOS ONE, 2021
Rapidly progressing antibiotic resistance is a great challenge in therapy. In particular, the infections caused by carbapenem-resistant Enterobacteriaceae (CRE) are exceedingly difficult to treat. Carbapenemase production is the predominant mechanism of resistance in CRE. Early and accurate identification of carbapenemase-producing carbapenem-resistant Enterobacteriaceae (CP-CRE) is extremely important for the treatment and prevention of such infections. In the present study, four phenotypic carbapenemase detection tests were compared and an algorithm was developed for rapid and cost-effective identification of CP-CRE. A total of 117 Enterobacteriaceae (54 CP-CRE, 3 non-CP-CRE, and 60 non-CRE) isolates were tested for carbapenemase production using modified Hodge test (MHT), modified carbapenem inactivation method (mCIM), Carba NP test (CNPt), and CNPt-direct test. The overall sensitivity/specificity values were 90.7%/92.1% for MHT, 100%/100% for mCIM, 75.9%/100% for CNPt, and 83.3%...
Scientific Reports
Rapid detection of carbapenemase-producing Enterobacteriaceae (CPE) represents a major challenge for microbiology laboratories. We evaluated the BYG Carba v2.0 using a simplified protocol, which detects CPE in less than 30 minutes. This new procedure reduces the hands-on-time from 5 to one minute and only requires a limited amount of material (one to three colonies) thereby preventing the need for subculturing bacterial isolates to reach a larger amount of pure biomass. This multicentre study involved four European reference laboratories. For the 1181 isolates tested across four centres, BYG Carba v2.0 yielded overall sensitivity and specificity of 96.3% (CI95: 94.5-97.5) and 99.7% (CI95: 98.6-100) respectively. Considering only the 670 consecutive isolates tested prospectively, the BYG Carba v2.0 displayed overall positive and negative predictive values of 99.7% (CI95: 95.4-98.9) and 97.5% (CI95: 94.9-98.8). Regarding time to positivity, 85% of CPE detected were positive within ten minutes. The BYG Carba v2.0 is a new highly simplified, rapid and accurate electrochemical assay discriminating between CPE and non-CPE in less than 30 min. The real-time quantified signal allows objective and traceable interpretation of the results. The emergence and worldwide spread of carbapenemase-producing Enterobacteriaceae (CPE) represents a major public health concern. Accurate and timely detection of CPE is essential for patient management and for rapid implementation of infection control measures 1, 2. Various confirmatory tests for the non-molecular detection of CPE have been proposed 3. The most rapid methods rely on the detection of carbapenem hydrolysis by colorimetric assay or by mass spectrometry 3. We recently evaluated three colorimetric assays, the RAPIDEC ® CARBA NP test (BioMérieux, Marcy l'Etoile,
Microbial drug resistance (Larchmont, N.Y.), 2016
Timely detection of carbapenemases by both phenotypic and genotypic methods is essential for developing strategies to control the spread of infections by carbapenem-resistant isolates and related morbidity and mortality. The aim of this study was to compare the performance of a commercial kit, the RAPIDEC(®) CARBA NP, and an in-house technique, the carbapenem inactivation method (CIM), against a panel of 136 carbapenemase- and noncarbapenemase-producing Enterobacteriaceae, Acinetobacter baumannii, and Pseudomonas aeruginosa isolates. RAPIDEC CARBA NP displayed 99% sensitivity and 100% specificity, whereas the sensitivity and specificity were 78% and 100% for the CIM test, respectively. A slight modification of the CIM test, a prolonged incubation time of 4 hours instead of two, increased the sensitivity of the test to 90% by diminishing false negativity particularly for A. baumannii. In conclusion, both tests possess a high performance and are practical for the detection of carbapen...
BMC Microbiology
Background Carbapenemase-resistant Enterobacteriaceae (CRE) cause many serious infections resulting in increasing treatment cost, prolonged hospitalization, and mortality rate. Reduced expression and/or mutations of porins and the presence of carbapenemase promote Enterobacteriaceae survival under carbapenem treatments. Development of accurate methods for the detection of antimicrobial resistance is required not only for therapy but also to monitor the spread of resistant bacteria or resistance genes throughout the hospital and community. In this study, we aimed to evaluate the phenotypic methods, Modified Hodge test (MHT), modified carbapenem inactivation method (mCIM), and EDTA-CIM (eCIM) for the detection of carbapenemase-producing Enterobacteriaceae (CPE). Results The results showed that mCIM had a sensitivity of 100% and a specificity of 100%, whereas the MHT had a sensitivity of 84.8% and a specificity of 97.8% for the 195 CRE isolates tested (105 CPE and 90 non-CPE isolates)....
Journal of clinical microbiology, 2017
The ability of clinical microbiology laboratories to reliably detect carbapenemase-producing carbapenem-resistant Enterobacteriaceae (CP-CRE) is an important element of the effort to prevent and contain the spread of these pathogens and an integral part of antimicrobial stewardship. Existing methods each have limitations. A new, straightforward, inexpensive, and specific phenotypic method for the detection of carbapenemase production, the carbapenem inactivation method (CIM), was recently described. Here we describe a two-stage evaluation of a modified carbapenem inactivation method (mCIM), in which tryptic soy broth was substituted for water during the inactivation step and the length of this incubation was extended. A validation study was performed in a single clinical laboratory to determine the accuracy of the mCIM, followed by a nine-laboratory study to verify the reproducibility of these results and define the zone size cut-off that best discriminated between CP-CRE and Entero...
JOURNAL OF CLINICAL AND DIAGNOSTIC RESEARCH, 2018
Introduction: Carbapenems have become one of the last resort of antimicrobials. But in last few years, Carbapenem Resistant Enterobacteriaceae (CRE) have been reported worldwide. Various phenotypic tests have been proposed for detection of carbapenemase activity including the newer modified Carbapenem Inactivation Method (mCIM) as advised by Clinical Laboratory Standard Institute (CLSI) 2017 guidelines. Aim: Detection of CRE from clinical specimens with new mCIM method and its comparative evaluation with phenotypic and genotypic methods. Materials and Methods: Study was conducted between January 2017 and December 2017 at KIMS, Karad. Total 66 CRE, isolated from 1634 clinical specimens and identified by VITEK 2 (Biomerieux, France) were included in the study. Phenotype screening was done by mCIM (CLSI 2017) method and was compared with Modified Hodge test (MHT) and Combined Disc Test (CDT) methods. Klebsiella pneumoniae ATCC BAA-1705 and Klebsiella pneumoniae ATCC BAA-1706 were used as positive and negative controls respectively. Molecular confirmation of these isolates for carbapenemase producing genes bla NDM-1 , bla OXA-48 , bla KPC was done by multiplex Polymerase Chain Reaction (PCR) study. Results: Klebsiella pneumoniae (n=35) outnumbered the other bacterial species among 66 CRE included in the study. mCIM was positive for 65 (98.48%) out of 66 isolates while MHT and CDT was positive for 50 (75.75%) and 59 (89.39%) of CRE isolates respectively. All the CRE isolates showed presence of at least one carbapenemase producing gene. Conclusion: The mCIM method is simple, less subjective, cost effective, reproducible and most sensitive method and plays important role in detection and prevention of spread of CRE, thereby, reducing morbidity and mortality, especially where there is lack of automation and molecular diagnostic facility.
Journal of Antimicrobial Chemotherapy, 2015
The objective of this study was the evaluation of the performance of two commercially available biochemical tests for the rapid detection of carbapenemase-producing Enterobacteriaceae compared with a home-made technique. Methods: A collection of 150 enterobacterial isolates, including 132 isolates with decreased susceptibility to at least one carbapenem molecule, were tested for carbapenemase activity using the RAPIDEC w CARBA NP (bioMérieux), the Rapid CARB Screen w (Rosco Diagnostica) and the home-made Carba NP test. This strain collection included 55 non-carbapenemase producers, 21 KPC producers, 21 NDM producers, 17 VIM producers, 11 IMP producers, 16 OXA-48 producers and 9 OXA-48-like producers (OXA-162, OXA-181, OXA-204, OXA-232 and OXA-244). Results: The RAPIDEC w CARBA NP detected all carbapenemase producers except a single OXA-244 producer. Using the Rapid CARB Screen w , one KPC-2, two NDM-1, one OXA-48 and five OXA-48 variant producers gave equivocal results and one OXA-244 producer was not detected. Using the Carba NP test, the same OXA-244 producer was not detected and one OXA-181 producer and one OXA-244 producer gave equivocal results. Sensitivity and specificity were 99% (95% CI 94.3%-99.8%) and 100% (95% CI 93.5%-100%), respectively, for the RAPIDEC w CARBA NP test, 89.5% (95% CI 81.7%-94.2%) and 70.9% (95% CI 57.9%-81.2%) for the Rapid CARB Screen w and 96.8% (95% CI 91.1%-98.9%) and 100% (95% CI 93.5%-100%) for the Carba NP test. The impact of the use of an adequate bacterial inoculum for obtaining the optimal performance with the RAPIDEC w CARBA NP was noted. Conclusions: The RAPIDEC w CARBA NP possesses the best performance for rapid and efficient detection of carbapenemase-producing Enterobacteriaceae.