CHROMagar™ ESBL/mSuperCARBA bi-plate medium for detection of ESBL- and carbapenemase-producing Enterobacteriaceae from spiked stools

doi:10.1016/j.diagmicrobio.2019.05.002

Diagnostic Microbiology and Infectious Disease

Volume 95, Issue 2, October 2019, Pages 107-112

https://doi.org/10.1016/j.diagmicrobio.2019.05.002 Get rights and content

Abstract

The recently released CHROMagar™ ESBL/CHROMagar™ mSuperCARBA bi-plate medium was evaluated for the detection of ESBL- and carbapenemase-producing Enterobacteriaceae. Spiked stools were used to mimic in vivo stool colonization. Two-hundred enterobacterial isolates were tested. Respective sensitivities of 93.9% and 97.8% were obtained for the detection of ESBL and carbapenemase producers.

Section snippets

Acknowledgments

We thank Saoussen Oueslati and Sandrine Bernabeu for helping constituting the panel of isolates.

Funding

This work was partially supported by the University Paris-Sud, France and Assistance Publique-Hôpitaux de Paris, Paris, France. LD, DG, and TN are members of the Laboratory of Excellence in Research on Medication and Innovative Therapeutics (LERMIT) supported by a grant from the French National Research Agency (ANR-10-LABX-33).

Transparency declaration

None to declare.

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Quantification of β-lactamase producing bacteria in German surface waters with subsequent MALDI-TOF MS-based identification and β-lactamase activity assay
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Environmental oligotrophic bacteria are suspected to be highly relevant carriers of antimicrobial resistance (AMR). However, there is a lack of validated methods for monitoring in the aquatic environment. Since extended-spectrum β-lactamases (ESBLs) play a particularly important role in the clinical sector, a culturing method based on R2A-medium spiked with different combinations of β-lactams was applied to quantify β-lactamase-producing environmental bacteria from surface waters. In German surface water samples (n = 28), oligotrophic bacteria ranging from 4.0 × 10³ to 1.7 × 10⁴ CFU per 100 mL were detected on the nutrient-poor medium spiked with 3^rd generation cephalosporins and carbapenems. These numbers were 3 log₁₀ higher compared to ESBL-producing Enterobacteriales of clinical relevance from the same water samples. A MALDI-TOF MS identification of the isolates demonstrated, that the method leads to the isolation of environmentally relevant strains with Pseudomonas, Flavobacterium, and Janthinobacterium being predominant β-lactam resistant genera. Subsequent micro-dilution antibiotic susceptibility tests (Micronaut-S test) confirmed the expression of β-lactamases. The qPCR analysis of surface waters DNA extracts showed the presence of β-lactamase genes (bla_TEM, bla_CMY-2, bla_OXA-48, bla_VIM-2, bla_SHV, and bla_NDM-1) at concentrations of 3.7 (±1.2) to 1.0 (±1.9) log₁₀ gene copies per 100 mL. Overall, the results demonstrate a widespread distribution of cephalosporinase and carbapenemase enzymes in oligotrophic environmental bacteria that have to be considered as a reservoir of ARGs and contribute to the spread of antibiotic resistance.
The role of the microbiology laboratory in the diagnosis of multidrug-resistant Gram-negative bacilli infections. Importance of the determination of resistance mechanisms
2022, Medicina Intensiva
Un diagnóstico y tratamiento precoz tiene un impacto importante en la morbimortalidad de las infecciones producidas por bacterias multirresistentes. Los bacilos gramnegativos multirresistentes constituyen la principal amenaza actual en los hospitales y muy especialmente en las unidades de cuidados intensivos. El papel del laboratorio de microbiología es esencial en dar una respuesta rápida y eficaz. En esta revisión se actualiza los procedimientos del laboratorio de microbiología para la detección rápida de los bacilos gramnegativos multirresistentes y de sus determinantes de resistencia. También se estudia el papel del laboratorio en la vigilancia y control de brotes por estas bacterias, incluyendo las técnicas de tipificación. Se destaca la importancia de proporcionar mapas de resistencia normalizados que permitan conocer la situación epidemiológica de las diferentes unidades. Finalmente se revisa la importancia de sistemas de comunicación eficaces para la transmisión de resultados y la toma de decisiones en el manejo de pacientes infectados por bacilos gramnegativos multirresistentes.
Early diagnosis and treatment has an important impact on the morbidity and mortality of infections caused by multidrug-resistant bacteria. Multidrug-resistant gram-negative bacilli constitute the main current threat in hospitals and especially in intensive care units. The role of the microbiology laboratory is essential in providing a rapid and effective response. This review updates the microbiology laboratory procedures for the rapid detection of multidrug-resistant gram-negative bacilli and its resistance determinants. The role of the laboratory in the surveillance and control of outbreaks caused by these bacteria, including typing techniques, is also studied. The importance of providing standardized resistance maps that allow knowing the epidemiological situation of the different units is emphasized. Finally, the importance of effective communication systems for the transmission of results and decision making in the management of patients infected by multidrug-resistant gram-negative bacilli is reviewed.
A European multicenter evaluation study to investigate the performance on commercially available selective agar plates for the detection of carbapenemase producing Enterobacteriaceae
2022, Journal of Microbiological Methods
Citation Excerpt :
As both these CPE-groups are the most common in human clinical settings and are the ones that primarily have been detected in food or animals (Bonardi and Pitino, 2019). Several studies have already been conducted on evaluating the performance of different CPE selective agar media (Cohen Stuart et al., 2013; Garcia-Fernandez et al., 2017; Girlich et al., 2013a; Girlich et al., 2013b; Girlich et al., 2019; Gottig et al., 2020). However, to our knowledge, no cross-country multicenter studies have been performed, nor a study including all commercially available selective agar plates on the market.
The European Food Safety Authority (EFSA) advised to prioritize monitoring carbapenemase producing Enterobacteriaceae (CPE) in food producing animals. Therefore, this study evaluated the performance of different commercially available selective agars for the detection of CPE using spiked pig caecal and turkey meat samples and the proposed EFSA cultivation protocol. Eleven laboratories from nine countries received eight samples (four caecal and four meat samples). For each matrix, three samples contained approximately 100 CFU/g CPE, and one sample lacked CPE. After overnight enrichment in buffered peptone water, broths were spread upon Brilliance™ CRE Agar (1), CHROMID® CARBA (2), CHROMagar™ mSuperCARBA™ (3), Chromatic™ CRE (4), CHROMID® OXA-48 (5) and Chromatic™ OXA-48 (6). From plates with suspected growth, one to three colonies were selected for species identification, confirmation of carbapenem resistance and detection of carbapenemase encoding genes, by methods available at participating laboratories. Of the eleven participating laboratories, seven reported species identification, susceptibility tests and genotyping on isolates from all selective agar plates. Agars 2, 4 and 5 performed best, with 100% sensitivity. For agar 3, a sensitivity of 96% was recorded, while agar 1 and 6 performed with 75% and 43% sensitivity, respectively. More background flora was noticed for turkey meat samples than pig caecal samples. Based on this limited set of samples, most commercially available agars performed adequately. The results indicate, however, that OXA-48-like and non-OXA-48-like producers perform very differently, and one should consider which CPE strains are of interest to culture when choosing agar type.
Metagenome and Resistome Analysis of Beta-Lactam-Resistant Bacteria Isolated from River Waters in Surabaya, Indonesia
2024, Microorganisms
Virulence, antimicrobial resistance, and molecular characteristics of carbapenem-resistant Klebsiella pneumoniae in a hospital in Shijiazhuang City from China
2023, International Microbiology
Rapid and automated screening of carbapenemase- and ESBL-producing Gram-negative bacteria from rectal swabs using chromogenic agar media and the ScanStation device
2023, Microbiology Spectrum

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Diagnostic Microbiology and Infectious Disease

Abstract

Section snippets

Acknowledgments

Funding

Transparency declaration

References (19)

Project management: importance for diagnostic laboratories

Clin Microbiol Infect

Laboratory automation in clinical bacteriology: what system to choose?

Clin Microbiol Infect

CHROMagar mSuperCARBA performance in carbapenem-resistant Enterobacteriaceae isolates characterized at molecular level and routine surveillance rectal swab specimens

Diagn Microbiol Infect Dis

CHROMagar mSuperCARBA and RAPIDEC^® Carba NP test for detection of carbapenemase-producing Enterobacteriaceae

Diagn Microbiol Infect Dis

Chromogenic culture media or rapid immunochromatographic test: which is better for detecting Klebsiella pneumoniae that produce OXA-48 and can they be used in blood and urine specimens

J Microbiol Methods

Automation in bacteriology: a changing way to perform clinical diagnosis in infectious diseases

Clin Microbiol Infect

Carbapenem resistance in Enterobacteriaceae: here is the storm!

Trends Mol Med

Extended-spectrum beta-lactamase-producing Enterobacteriaceae: an emerging public-health concern

Lancet Infect Dis

Review: Molecular detection of extended spectrum-beta-lactamase– and carbapenemase-producing Enterobacteriaceae in a clinical setting

Diagn Microbiol Infect Dis

Cited by (13)

Quantification of β-lactamase producing bacteria in German surface waters with subsequent MALDI-TOF MS-based identification and β-lactamase activity assay

The role of the microbiology laboratory in the diagnosis of multidrug-resistant Gram-negative bacilli infections. Importance of the determination of resistance mechanisms

A European multicenter evaluation study to investigate the performance on commercially available selective agar plates for the detection of carbapenemase producing Enterobacteriaceae

Metagenome and Resistome Analysis of Beta-Lactam-Resistant Bacteria Isolated from River Waters in Surabaya, Indonesia

Virulence, antimicrobial resistance, and molecular characteristics of carbapenem-resistant Klebsiella pneumoniae in a hospital in Shijiazhuang City from China

Rapid and automated screening of carbapenemase- and ESBL-producing Gram-negative bacteria from rectal swabs using chromogenic agar media and the ScanStation device

BacteriologyCHROMagar™ ESBL/mSuperCARBA bi-plate medium for detection of ESBL- and carbapenemase-producing Enterobacteriaceae from spiked stools

Abstract

Section snippets

Acknowledgments

Funding

Transparency declaration

Clin Microbiol Infect

Clin Microbiol Infect

Diagn Microbiol Infect Dis

Diagn Microbiol Infect Dis

J Microbiol Methods

Clin Microbiol Infect

Trends Mol Med

Lancet Infect Dis

Diagn Microbiol Infect Dis

Bacteriology
CHROMagar™ ESBL/mSuperCARBA bi-plate medium for detection of ESBL- and carbapenemase-producing Enterobacteriaceae from spiked stools