ChromID® CARBA Agar Fails to Detect Carbapenem-Resistant Enterobacteriaceae With Slightly Reduced Susceptibility to Carbapenems

Pauly, Natalie; Hammerl, Jens A.; Grobbel, Mirjam; Tenhagen, Bernd‐Alois; Käsbohrer, Annemarie; Bisenius, Sandra; Fuchs, Jannika; Horlacher, Sabine; Lingstädt, Holger; Mauermann, Ute; Mitro, Silke; Müller, Melanie; Rohrmann, Stefan; Schiffmann, Arthur; Stührenberg, B.; Zimmermann, Pia; Schwarz, Štefan; Meemken, Diana; Irrgang, Alexandra

doi:10.3389/fmicb.2020.01678

Cited by 8 publications

(15 citation statements)

References 55 publications

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“…Further, we exchanged the commercial selective agar chromID ® CARBA agar with an in-house prepared McC + CTX and McC + CTX + MEM. Our results support a previous study on the suitability of chromID ® CARBA agar for detecting CPE with only slightly reduced susceptibility to carbapenems [22]. Within this study, it is presumed that the concentration of different ingredients and thus the selectivity is too high for CPE originating from animals or food.…”

Section: Modification Steps Of the Official Isolation Protocolsupporting

confidence: 89%

“…After estimating bacterial survival, other options for improved detection of the target bacteria were tested. To account for the low MIC values of CPE isolated from samples of animal origin [22], we considered triggering gene expression by using a sublethal concentration of MEM. Moreover, the usage of ZnSO 4 to push the metallo-β-lactamases was tested.…”

Section: Modification Steps Of the Official Isolation Protocolmentioning

confidence: 99%

“…Within this study, it is presumed that the concentration of different ingredients and thus the selectivity is too high for CPE originating from animals or food. CPE from livestock often exhibit lower MIC values for the respective carbapenems than clinical human isolates [22,24]. Our previous study investigated the use of chromID ® CARBA after the end of the shelf life expecting a possible decrease in antimicrobial concentration.…”

Section: Modification Steps Of the Official Isolation Protocolmentioning

confidence: 99%

“…Several studies described the chromID ® CARBA agar as one of the most sensitive and specific chromogenic media for the detection of CP Enterobacteriaceae (CPE) in clinical samples in human medicine [18][19][20][21]. In Germany, this agar represents one of the most commonly used selective media for CPE monitoring purposes [22]. By using a highly sensitive molecular pre-screening of enrichment cultures (i.e., by real-time PCR), laborious microbiological investigations might be avoided, and further efforts can focus on the recovery of CPE from presumptive-positive samples.…”

Section: Introductionmentioning

confidence: 99%

“…Thus, we identified the necessity to improve the isolation method for CP E. coli from livestock samples, especially from caecum content. Difficulties in the microbiological processing of caeca samples are the growth of an accompanying microbiota, the low CPE concentrations, and the considerably lower MIC values of CPE from animal origin in comparison to human isolates [22]. Here, we present comparative analyses on the performance of the EURL-method with a number of alterations potentially leading to an improvement of the CPE detection and recovery rate.…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Isolation Procedure for CP E. coli from Caeca Samples under Review towards an Increased Sensitivity

Pauly

Klaar

Skladnikiewicz-Ziemer

et al. 2021

Microorganisms

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Due to the increasing reports of carbapenemase-producing Enterobacteriaceae (CPE) from livestock in recent years, the European Reference Laboratory for Antimicrobial Resistances (EURL-AR) provided a protocol for their recovery from caecum and meat samples. This procedure exhibited limitations for the detection of CPE with low carbapenem MIC values. Therefore, it was modified by a second, selective enrichment in lysogeny broth with cefotaxime (CTX 1 mg/L) and with meropenem (MEM 0.125 mg/L) at 37 °C under microaerophilic conditions. By Real-time PCR, these enrichments are pre-screened for the most common carbapenemase genes. Another adaptation was the use of in-house prepared MacConkey agar with MEM and MEM+CTX instead of commercial selective agar. According to the EURL-method, we achieved 100% sensitivity and specificity using the in-house media instead of commercial agar, which decreased the sensitivity to ~75%. Comparing the method with and without the second enrichment, no substantial influence on sensitivity and specificity was detected. Nevertheless, this enrichment has simplified the CPE-isolation regarding the accompanying microbiota and the separation of putative colonies. In conclusion, the sensitivity of the method can be increased with slight modifications.

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Section: Modification Steps Of the Official Isolation Protocolsupporting

confidence: 89%

Section: Modification Steps Of the Official Isolation Protocolmentioning

confidence: 99%

Section: Modification Steps Of the Official Isolation Protocolmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Isolation Procedure for CP E. coli from Caeca Samples under Review towards an Increased Sensitivity

Pauly

Klaar

Skladnikiewicz-Ziemer

et al. 2021

Microorganisms

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

Dierikx

Börjesson

Perrin-Guyomard

et al. 2022

Journal of Microbiological Methods

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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.

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Infiltration of hidden antimicrobial resistance among healthy people in a Japanese community

Fukuda

Nakamura

Umeda

et al. 2022

JAC-Antimicrobial Resistance

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Background Under non-antimicrobial selective pressure, antimicrobial-resistant bacteria do not easily become dominant in the microbiota. Furthermore, their low levels prevent detection by isolation, resulting in an underestimation of the prevalence of antimicrobial-resistant bacteria. Objectives We evaluated the infiltration of antimicrobial-resistant bacteria and their related β-lactamase genes among healthy people in non-clinical settings. Methods Cephalosporin- and fluoroquinolone-resistant Escherichia coli and bla genes were quantified in 217 faecal samples from healthy people in non-clinical settings in Japan. E. coli colonies grown on deoxycholate hydrogen sulphide-lactose (DHL) agar, with and without antimicrobials (cefotaxime and ciprofloxacin), were quantified, and E. coli isolates were analysed for their susceptibility to antimicrobials and the presence of bla genes. DNA extracted from faecal samples was used to quantify bla genes using quantitative PCR (qPCR). Results The isolation rates of cefotaxime- and ciprofloxacin-resistant E. coli were 6.9% and 12.4%, respectively, using agars without antimicrobials, and 12.0% and 24.4%, respectively, using agars with antimicrobials. For samples from which cefotaxime- and ciprofloxacin-resistant E. coli were isolated only using agars with antimicrobials, the ratios of cfu on DHL agars with and without antimicrobials were below −2 log. E. coli harbouring bla genes were isolated from 35.0% of the faecal samples using agars, and bla genes were detected in 65.0% of faecal DNA samples using qPCR. Conclusions Among people carrying cefotaxime- and ciprofloxacin-resistant E. coli in non-clinical settings, cefotaxime- and ciprofloxacin-resistant E. coli were not dominant in half of the subjects. These individuals may play a role as reservoirs of antimicrobial-resistant bacteria.

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ChromID® CARBA Agar Fails to Detect Carbapenem-Resistant Enterobacteriaceae With Slightly Reduced Susceptibility to Carbapenems

Cited by 8 publications

References 55 publications

Isolation Procedure for CP E. coli from Caeca Samples under Review towards an Increased Sensitivity

Isolation Procedure for CP E. coli from Caeca Samples under Review towards an Increased Sensitivity

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

Infiltration of hidden antimicrobial resistance among healthy people in a Japanese community

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