The samples were collected in the newly opened intensive care unit (10 stations) where COVID-19 patients were hospitalized. Samples were collected using Amies media transport swabs (Copan). Rectal swabs for carbapenemase-producing bacilli were inoculated on chromID CARBA SMART Agar (bioMerieux), chromID ESBL Agar (bioMerieux), chromID VRE Agar (bioMerieux), chromID MRSA SMART (bioMerieux) and incubated aerobically at 35-37°C for 24h. Other clinical samples (urine, BAL, nasal/throat swab, blood culture) were inoculated on appropriate culture media and incubated aerobically at 35-37°C for 24h. Swabs from the hospital environment, after 24h pre-incubation at 35-37°C, were inoculated on the following culture media: Columbia Agar (bioMerieux), Mac Conkey Agar (bioMerieux), Sabouraud Gentamicin Chloramphenicol 2 Agar (bioMerieux), chromID CARBA SMART Agar (bioMerieux), chromID ESBL Agar (bioMerieux), chromID VRE Agar, chromID MRSA SMART and incubated for another 24h under aerobic conditions at 35-37°C. Bacterial colonies were identified using VITEK2 cards, and the resistance mechanisms were determined using the disk diffusion method. An OKNVI RESIST-5 (CORIS BioConcept) immunochromatographic tests were performed from the strains insensitive to carbapenems, enabling the determination of the type of carbapenemase (KPC, NDM VIM, OXA-48, OXA-163).
Chromid vre agar
Manufactured by bioMérieux
Sourced in France, Germany
ChromID VRE agar is a chromogenic culture medium used for the detection and differentiation of vancomycin-resistant Enterococcus (VRE) from clinical specimens. The agar contains chromogenic substrates that allow the identification of VRE based on the colony color.
Automatically generated - may contain errors
Lab products found in correlation
Vitek 2, by bioMérieux (3 mentions)
Chromid esbl agar, by bioMérieux (3 mentions)
Chromid mrsa smart, by bioMérieux (3 mentions)
Chromid carba smart agar, by bioMérieux (2 mentions)
Etest, by bioMérieux (2 mentions)
Maldi tof, by Bruker (2 mentions)
Maldi tof ms, by Bruker (1 mentions)
Amies transport media, by bioMérieux (1 mentions)
Chromid mrsa agar, by bioMérieux (1 mentions)
Macconkey agar, by bioMérieux (1 mentions)
Sabouraud gentamicin chloramphenicol 2 agar, by bioMérieux (1 mentions)
Columbia agar, by bioMérieux (1 mentions)
Bile esculin agar, by HiMedia (1 mentions)
Brain heart infusion, by HiMedia (1 mentions)
Vitek 2 system, by bioMérieux (1 mentions)
Brilliance vre agar, by Thermo Fisher Scientific (1 mentions)
Columbia blood agar cba, by Thermo Fisher Scientific (1 mentions)
Amies collection and transport medium, by Hain Lifescience (1 mentions)
Vitek ms, by bioMérieux (1 mentions)
Vitek 2 instrument, by bioMérieux (1 mentions)
14 protocols using chromid vre agar
1
Screening COVID-19 Patients for Multidrug-Resistant Bacteria
2
Screening for ESBL and VRE Bacteria
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Rectal swabs were directly plated on chromID® ESBL agar (bioMérieux) for ESBL screening and enriched EnterococcoselTM Broth (Bile Esculin Azide Broth) (BD; Becton, Dickinson and Company) was used for VRE screening and subsequently cultured on chromID® VRE agar (bioMérieux).
Species identification and antibiotic susceptibility testing was done by MALDI-TOF MS (Bruker Daltonik GmbH, Bremen) and VITEK®2 (bioMérieux), respectively, following EUCAST criteria. Confirmation of ESBL was performed using disk diffusion (cefotaxime 30 μg, cefotaxime 30 μg plus clavulanic acid 10 μg, ceftazidime 30 μg, ceftazidime plus clavulanic acid 10 μg, cefepime 30 μg, cefepime 30 μg plus clavulanic acid 10 μg, and cefoxitin 30 μg) (Mast Diagnostics, Bootle, UK).
Species identification and antibiotic susceptibility testing was done by MALDI-TOF MS (Bruker Daltonik GmbH, Bremen) and VITEK®2 (bioMérieux), respectively, following EUCAST criteria. Confirmation of ESBL was performed using disk diffusion (cefotaxime 30 μg, cefotaxime 30 μg plus clavulanic acid 10 μg, ceftazidime 30 μg, ceftazidime plus clavulanic acid 10 μg, cefepime 30 μg, cefepime 30 μg plus clavulanic acid 10 μg, and cefoxitin 30 μg) (Mast Diagnostics, Bootle, UK).
3
Screening for Carbapenem-Resistant Bacteria
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Rectal swabs for carbapenemase-producing bacilli were taken and placed in Amies transport media (bioMérieux, France); inoculated on CHROMID® CARBA SMART agar (bioMérieux, France), CHROMID® ESBL agar (bioMérieux, France), CHROMID® VRE agar (bioMérieux, France), and CHROMID® MRSA SMART (bioMérieux, France); and incubated aerobically at 35–37°C for 24 h.
4
Screening for Antimicrobial-Resistant Bacteria
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Rectal swabs were cultured on chromogenic agar for detection of ESBL-producing Enterobacteriaceae and vancomycin resistant enterococcci (ID™ ESBL agar and chrom ID™ VRE agar (bioMérieux, Marcy l’Etoile, France)). The Centers for Disease Control and Prevention (CDC) method was applied to detect CPE [32 ,33 ]. Nasal swabs were cultured on chrom ID™ MRSA agar (bioMérieux, Marcy l’Etoile, France). In each case suspect colonies were subcultured for identification by standard methods and susceptibility testing was performed in accordance with EUCAST disk diffusion methods [34 ].
5
Rapid Detection of Vancomycin-Resistant Enterococci
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Stool samples positive for the GDH antigen were cultured in brain heart infusion (Himedia, India) with the supplement of gram-positive non-spore-forming bacteria (Himedia, India) under aerobic conditions for 48 h at 37 °C. The samples for VRE identification were grown on CHROMID® VRE agar (bioMérieux, France). The plates were grown in aerobic conditions for 24 h at 37 °C. Samples with negative results after cultivation on CHROMID® VRE were further tested for vancomycin-sensitive enterococci (VSE) isolates. The samples were grown on Bile esculin agar (Himedia, Czech Republic) with vancomycin 30-µg disk overnight at 37 °C.
6
Rapid Vancomycin-Resistant Enterococcus Detection
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Specimens were plated directly onto ChromID VRE agar (bioMérieux, Marcy l'Etoile, France). After 24–48 hours of incubation, any purple (Enterococcus faecium) or blue (Enterococcus faecalis) colonies were subcultured onto a non-selective medium for antimicrobial susceptibility testing using the Vitek 2 AST P602 card in the Vitek 2 system (bioMérieux, Hazelwood, MO). Negative results were not reported until 48 hours. Microorganisms used as controls were E. faecalis ATCC29212, QC-06A E. faecalis ATCC51299, and vancomycin-resistant E. faecium. According to the package insert, the sensitivity and specificity of ChromID VRE agar at day 2 without enrichment is 97.4% and 95.6%, respectively.
7
Vancomycin-resistant Enterococcus characterization
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A collection of previously characterized isolates of E. faecium (n = 13), Enterococcus faecalis (n = 11), Enterococcus gallinarum, Enterococcus casseliflavus, Enterococcus raffinosus and Enterococcus hirae was selected for testing. The isolates originated from NCTC collections (n = 12), a retrospective collection of bacteremia isolates from Cambridge University Hospitals (CUH) NHS Foundation Trust (2006–2012, n = 15), and from a meat sample (n = 1) (Table 1 ). They included isolates with vanA and vanB acquired resistance, and in the case of E. faecalis and E. faecium bacteremia isolates, a range of diverse hospital-adapted STs Raven et al., 2016 (link), Raven et al., 2016 ). The organisms were grown from glycerol bead vials stored at −80°C onto Columbia blood agar (CBA, Oxoid) and 200 μl of a 0.5 McFarland suspension was streaked using the four-quadrant technique onto chromID VRE agar (bioMérieux) and Brilliance VRE agar (Oxoid). Growth was assessed after 24 and 48 hours aerobic incubation at 37 °C. Vancomycin minimum inhibitory concentration was measured using Etest (bioMérieux).
8
Screening and Identification of MDROs
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For MDRO screening culture swabs were transferred from Amies collection and transport medium (Hain Lifescience, Nehren, Germany) onto selective CHROMagar ESBL plates (Mast Diagnostica, Paris, France). Matrix-assisted laser desorption ionization-time of flight analysis (VITEK MS, bioMérieux, Nürtingen, Germany) was used for identification of bacteria isolates, followed by performance of an antibiogram and resistogram using VITEK 2 and/or antibiotic gradient tests (bioMérieux). Carbapenemases detection was performed via PCR analysis and subsequent sequencing from carbapenem-resistant Enterobacteriaceae including the bla genes for carbapenemases NDM, VIM, IMP, OXA-48, OXA-48 like, and KPC as well as OXA-23, OXA-24, OXA-51, and OXA-58 for Acinetobacter baumannii [9 (link), 10 (link)]. For VRE detection swabs from Amies collection and transport medium (Hain Lifescience, Nehren, Germany) were transferred onto ChromID VRE agar (bioMérieux).
9
Vancomycin-Resistant Enterococcus Screening
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Rectal swabs were directly inoculated onto a chromogenic agar plate (ChromID VRE agar, bioMérieux, Marcy I'Etoile, France) containing 8 µg/mL vancomycin and incubated aerobically at 35℃. The agar plates were screened for growth of presumptive colonies after 24 hr and 48 hr of incubation. For colonies resembling enterococci, gram staining and the pyrrolidonyl arylamidase (PYR) test were performed. Then, gram-positive, PYR-positive cocci were screened for vancomycin resistance. Colonies were inoculated onto blood agar plates containing a 30-µg vancomycin disk. Following an incubation period of 24 hr, colonies that demonstrated vancomycin resistance were submitted for identification and antimicrobial susceptibility testing.
10
Routine VRE Screening Protocol
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Routine VRE screening at University Medical Centre Hamburg-Eppendorf, Germany, consists of a two-tier approach combining culture-based and molecular techniques (Fig. 1 b). Briefly, 100 µl of Amies medium from the rectal swabs (eSwab, Copan) were transferred to 2 ml of VRE enrichment broth (Oxoid, Basinstoke, UK) and incubated over-night. Enriched broth was plated on ChromID VRE agar (bioMérieux, Marcy l’Etoile, France) and incubated at 37 °C for 24 to 48 h. After colonies displaying morphology typical of VRE on the VRE agar was detected, nucleic acids were extracted using the MagNA-pure96 system (Roche) with 200 µl extraction volume according to manufacturer’s recommendation and further analysed by qPCR on the LightCycler 480 II (Roche) using vanA and vanB specific primers and probes24 (link). Simultaneously, colonies were identified by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF, Bruker, Billerica, MA, U.S.A.) and automated susceptibility testing on a Vitek2 instrument (Biomérieux, Marcy-l’Etoile, France).
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