Chromid candida

Manufactured by bioMérieux

Sourced in France

ChromID Candida is a chromogenic culture medium used for the detection and identification of Candida species in clinical samples. It is designed to facilitate the rapid isolation and differentiation of the most clinically relevant Candida species.

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CHROMID® Candida agar

8 protocols using chromid candida

Microbiological Identification Protocol

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Microbiological tests were carried out using classical methods routinely used in microbiological diagnostics. The material collected from the examined patients was cultured on appropriate culture media to multiply and then isolate pure microbial cultures. Aerobic bacteria were multiplied on solid medium Columbia agar with a 5% addition of sheep blood at 37°C. Anaerobic bacteria were multiplied on Schaedler K3 solid medium with a 5% addition of sheep blood at 37°C in an-aerobic conditions obtained using Biomerieux Genbag Anaer kits (Marcy l’Etoile, France). Candida fungi were multiplied and initially identified using chromogenic medium ChromID Candida from Biomerieux (Marcy l’Etoile, France).
After isolation and multiplication of the cultured strains of microorganisms, their species identification was carried out using the following sets of reagents: ENTEROtest 24 N, NEFERMtest 24 N, STREPTOtest 24, STAPHYtest 24, ANAEROtest 23, OXItest, PYRAtest, and the computer program TNW_lite 6.5 for species identification of Erba-Lachema microorganisms (Brno, Czech Republic). The following biochemical tests from Biomerieux (Marcy l’Etoile, France) were also used: Catalysis, Slidex Staph Kit, and API Candida. Execution, reading, and interpretation of the test results were performed in accordance with the recommendations of manufacturers of diagnostic reagent kits.

Pachoński M., Koczor-Rozmus A., Mocny-Pachońska K., Łanowy P., Mertas A, & Jarosz-Chobot P. (2021). Oral microbiota in children with type 1 diabetes mellitus. Pediatric Endocrinology, Diabetes, and Metabolism, 27(2), 100-108.

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Comprehensive Oral Microbial Profiling

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Oral microbiological examination was performed by taking a swab from the bottom of the mouth, using sterile swabs, in tubes with AMIES transport medium, with charcoal (DELTALAB, Rubi, Spain). The following media from Biomerieux (Marcy l’Etoile, France) were used to cultivate microorganisms: Columbia agar, Schaedler K3, ChromID Candida and Genbag anaer kits for the cultivation of anaerobic bacteria. Species identification of the cultured microorganisms was carried out using the following sets of reagents: ENTEROtest 24N, NEFERMtest 24N, STREPTOtest 24, STAPHYtest, ANAEROtest 23, OXItest, PYRAtest and the computer program TNW lite 6.5 for the identification of microorganisms by Erba-Lachema (Brno, Czech Republic). Biochemical tests by Biomerieux (Mercy l’ Etoile, France) were also used: Katalase, Slidex Staph Kit, and API Candida.

Wyszyńska M., Czelakowska A., Rosak P., Kasperski J., Łopacińska M., Ghanem A., Mertas A, & Skucha-Nowak M. (2023). The Impact of COVID-19 on the Oral Bacterial Flora in Patients Wearing Complete Dentures and on the Level of Exhaled Nitric Oxide as a Marker of Inflammation. Journal of Clinical Medicine, 12(17), 5556.

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Identification of Candida species

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All clinical isolates were first plated on Sabouraud dextrose agar (SDA, Thermo Fisher Oxoid, UK) for 24 h at 37 °C. Afterwards, germ-tube test was performed to differentiate between Candida spp. Colonies of the cultivated yeasts were suspended in sterile saline and adjusted to the density of McFarland 2. For the morphological examination, human serum was used. Two drops of the yeast suspension were applied to wells of microtiter plates with 30 µL of human serum. After incubation for 2–3 h at 37 °C, the number of the germ tube and the total number of the cells was calculated in Bürker counting chamber. Production of chlamydospores, blastospores, true hyphae, and pseudohyphae was compared.
Parallelly, CHROMID^® Candida (bioMérieux^®, Craponne, France) was used for presumptive identification of Candida strains, based on the pigmentation of the developing colonies due to different enzyme activity. The results were interpreted after 48 h of incubation at 30 °C, according to colony’s color and manufacturer’s instructions. Subsequently, for identification of the medically important isolated yeasts, the colorimetric sugar assimilation test Auxacolor 2 (Bio-Rad^®, Hercules, CA, USA) served for accurate identification. The method was applied as described in the kit’s guideline and results were then evaluated.

Černáková L., Líšková A., Lengyelová L, & Rodrigues C.F. (2022). Prevalence and Antifungal Susceptibility Profile of Oral Candida spp. Isolates from a Hospital in Slovakia. Medicina, 58(5), 576.

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Candida Identification Protocol

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The available samples were re-cultured on a chromID® Candida and incubated aerobically at 37 °C for 72 h. After visual inspection, one colony of each phenotypical type from each sample was selected for further analysis, re-cultured aerobically for 24 h at 37 °C on a Sabouraud Glucose agar (bioMérieux, France), and prepared for biochemical identification by the VITEK®2 (bioMérieux, France) system, according to the manufacturer’s manual.

Monsen R.E., Kristoffersen A.K., Gay C.L., Herlofson B.B., Fjeld K.G., Hove L.H., Nordgarden H., Tollisen A., Lerdal A, & Enersen M. (2023). Identification and susceptibility testing of oral candidiasis in advanced cancer patients. BMC Oral Health, 23, 223.

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Identification of Candida Species via RFLP-PCR

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All isolates were plated onto Sabouraud dextrose agar (Difco, USA) and incubated at 37 °C for 24 h. Presumptive identification was performed considering colony morphology and color on ChromID Candida (bioMérieux, France) and Candida chromogenic agar (CONDA, Spain) agars and subsequently confirmed using the API 32C (bioMérieux) auxanogram according to manufacturers. DNA was extracted using the UltraClean® Microbial DNA Isolation Kit (MoBio, USA) following the recommendations of the manufacturers. Definitive identification was reached either by amplification of a short portion of the SADH gene using a conventional RFLP-PCR protocol, as previously described [1 (link), 9 (link)] or by ITS sequencing when the obtained RFLP-PCR profile from the former technique was inconclusive.

Trobajo-Sanmartín C., Ezpeleta G., Pais C., Eraso E, & Quindós G. (2018). Design and validation of a multiplex PCR protocol for microsatellite typing of Candida parapsilosis sensu stricto isolates. BMC Genomics, 19, 718.

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Characterization of Candida albicans Isolates

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Three strains used as control strains were as follows: Ca11r—C. albicans (ATCC 10231) and Ca12r—C. albicans (ATCC 60193) and Ca1r—clinical C. albicans strain identified by MALDI. Seven clinical isolates came from vaginal candidiasis patients (Ca1V–Ca7V) and have been kindly provided by Dr. Agata Karowicz-Bilińska (Medical University of Lodz). 28 strains (Ca1s–Ca82s) were isolated from the saliva of healthy volunteers (Bioethics Committee agreement no. 6./KBBN-UŁ/II/2014). Clinical strains were isolated on yeast extract–peptone–dextrose (YEPD) agar plates supplemented with chloramphenicol. Then, they were directly identified as C. albicans species using chromogenic culture media (chromID^®Candida, bioMérieux), HRM and sequence analysis of ITS2 regions (data not shown). All strains were stored at −70 °C as stocks in 8 % DMSO. Strains were cultured for 24 h at 37 °C in YEPD-agar containing 2 % glucose, 2 % peptone, and 1 % yeast extract.

Caban M., Strapagiel D., Dziadek J., Korycka-Machała M, & Grzelak A. (2016). Principles of a New Protocol for Prediction of Azole Resistance in Candida albicans Infections on the Basis of ERG11 Polymorphisms. Current Microbiology, 73, 172-182.

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Diagnosis and Antifungal Susceptibility of COVID-19 ICU Patients

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Three sets of blood cultures were drawn in Bactalert FA Plus (bioMerieux, Marcy-l’Étoile, France) at admission from every critical COVID-19 patient transferred to the ICU from the isolation ward. Blood cultures were also retaken if the patient developed clinical signs of sepsis, had persistent fevers or deteriorated clinically, without other plausible explanations. After microscopic examination, fungal isolation was achieved by culturing positive blood cultures on Candida chromagar (CHROMID Candida, bioMérieux) and Sabouraud-CAF agar (Sabouraud Gentamicin Chloramphenicol 2, bioMérieux). Fungal identification was performed by a matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) system (Bruker Daltonik GmbH, Bremen, Germany). A broth microdilution system (Sensititre Yeast Oneplates, ThermoFisher Scientific, Waltham, Massachusetts, US) was used for testing fungal susceptibilities for amphotericin-B, azoles and echinocandins, following manufacturer’s instructions. The guidelines of the European Committee on Antimicrobial Susceptibility Testing (EUCAST) were used for the interpretations of fungal isolate susceptibility [18 ].

Alessandri F., Ceccarelli G., Migliara G., Baccolini V., Russo A., Marzuillo C., Ceparano M., Giordano G., Tozzi P., Galardo G., Raponi G., Mastroianni C., Venditti M., Pugliese F, & d’Ettorre G. (2023). High Incidence of Candidemia in Critically Ill COVID-19 Patients Supported by Veno-Venous Extracorporeal Membrane Oxygenation: A Retrospective Study. Journal of Fungi, 9(1), 119.

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Oral Tumor Microbiome Profiling

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Oral swabs were done from the surface of the tumor. Bacterial cultures were performed in the Laboratory of Bacteriology of Medical Diagnostic Laboratory of Fryderyk Chopin Provincial Specialist Hospital No. 1 in Rzeszów, Poland. Columbia Agar with 5% sheep blood, CHROMID® CPS® Elite, CHROMID® MRSA, and CHROMID® Candida (all of them bioMérieux SA, France) solid media were used for identification of aerobic bacteria and candida. Cultures for identification of bacteria were incubated for 24 h at 37°C and for candida culture for 48 h at 35°C.For isolation of anaerobic bacteria Schaedler agar +5% sheep blood (bioMérieux SA, France) were used and was incubated for 48 h in GENBAG ANAEROBIC sachets (bioMérieux SA, France) in an incubator at 37°C. Bacteria and yeasts were identified with the use of VITEK® 2 cards (bioMérieux SA, France) according to the manufacturer's protocol.

Zielińska K., Karczmarek-Borowska B., Kwaśniak K., Czarnik-Kwaśniak J., Ludwin A., Lewandowski B, & Tabarkiewicz J. (2020). Salivary IL-17A, IL-17F, and TNF-α Are Associated with Disease Advancement in Patients with Oral and Oropharyngeal Cancer. Journal of Immunology Research, 2020, 3928504.

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