Chromogenic Culture Media: Principle, Composition, Results

Chromogenic culture media are used to isolate, identify, and differentiate specific microorganisms from a heterogeneous population. The medium contains chromogenic substrate which is utilized by the microorganisms to give colored colonies that is specific for each microorganism. Depending on the color of the result, the presence or absence of the target organism is determined and also accurately differentiated from others.

Classical culture media are based on the principle of change in color of the pH indicator whereas Chromogenic media are based on enzymatic utilization of chromogenic substrates.

Principle of Chromogenic Media

Chromogenic media contains soluble colorless molecules called chromogens. Chromogens are composed of two parts: a substrate (which is the target of specific enzymatic activity of the microorganism) and a chromophore.

Principle of Chromogenic Agar Medium (source: chromagar.com)

When the bond between the substrate and chromophore is split by a specific enzyme produced by the target microorganism, chromophore is released. In its unconjugated form, the chromophore shows distinctive color. Due to reduced solubility, chromophore forms a precipitate that imparts unique color to the colony.

Composition of Chromogenic Agar Media

Like traditional culture media, chromogenic media also contains nutrients such as peptones, amino acids, yeast extract, minerals, vitamins, and solidifiers (agar). Depending on the purpose chromogenic media may also contain inhibitors. Unlike traditional media, they contain chromogenic substrates or chromogens. These chromogenic substrates such as ONPG, X-Gal, or X-Glu, together with a specified selectivity of the medium, is the simple principle behind chromogenic media.

Chromogenic Agar Media (Source:biomerieux)

Advantages of Use of Chromogenic Agar Media:

  1. Less Labor intensive and more economic:
    Though people perceive chromogenic media as an expensive alternative, the use of a single chromogenic medium rather two-three selective ones reduces the cost of sample processing. Chromogenic media may eliminate the need for subculture and further biochemical tests for the identification of the isolates. 
  2. Less time consuming:  As chromogenic media eliminate various steps of sample processing (e.g. subculturing, biochemical testing) results are available within 24 hours as compared to 48 hours or more by conventional methods.  Timely diagnosis not only ensures a better outcome for the patients but also helps in the prevention and spread of infections.
  3. Easy identification: On chromogenic agar medium, target colonies of specific microorganisms can be recognized by their color at a glance. No specialized equipment needed.

Examples of Chromogenic Agar Medium

Most of the commercially available chromogenic media utilize substrates like indoxyl and its halogenated derivatives, metal chelators like Esculin, 8‐Hydroxyquinoline, substrates for β‐glucosidase enzyme etc. Based on these a number of companies have formulated several variations depending on need and usage.

Following are some of the examples

S.NTarget organismMedia Available
1.Candida albicansCHROMagar Candida, Albicans ID, CandiSelect, Candida ID, Pourmedia Vi Candida, Chromogenic Candida agar , Brilliance Candida Agar  and HiCrome Candida differential agar. 
2.Pseudomonas aeruginosaChrom ID Pseudomonas 
3.Urinary Tract pathogensCHROMagar Orientation, UriSelect medium, Rainbow Agar UTI medium, Chromogenic UTI medium, USA agar, Harlequin CLED and Urichrom agar 
4.Bacillus cereusBacillus cereus ChromoSelect Agar 
5.Escherichia coli serotype O157:H7 CHROMagar O157
6.Methicillin resistant Staphylococcus aureus (MRSA)CHROMagar MRSA II 
7.Carbapenem-resistant EnterobacteriaceaeBrilliance CRE Agar

References

  1. Perry J. D. (2017). A Decade of Development of Chromogenic Culture Media for Clinical Microbiology in an Era of Molecular Diagnostics. Clinical microbiology reviews, 30(2), 449–479. https://doi.org/10.1128/CMR.00097-16 
  2. Garcia, B. L. N., Fidelis, C. E., Freu, G., Granja, B. M., & Dos Santos, M. V. (2021). Evaluation of Chromogenic Culture Media for Rapid Identification of Gram-Positive Bacteria Causing Mastitis. Frontiers in veterinary science, 8, 662201. https://doi.org/10.3389/fvets.2021.662201

Nisha Rijal

I am working as Microbiologist in National Public Health Laboratory (NPHL), government national reference laboratory under the Department of health services (DoHS), Nepal. Key areas of my work lies in Bacteriology, especially in Antimicrobial resistance.

5 thoughts on “Chromogenic Culture Media: Principle, Composition, Results

  1. Iam a medical laboratory Technician , working in national Public Health laboratory (NPHL)
    my question is , what are the most challenges in antibiotic resistance to Gram negative Bacteria ?

  2. I am writing a proposal to do a research on carbapenem resistant enterobacteriaceae, so ho can I get a brilliance CRE Agar?

  3. I am a microbiologist at a National Reference laboratory in Ethiopia. All my works are on bacterial diseases and antimicrobial resistance. Currently, I am working on food borne illnesses. My question is what is the specific components of ChromSTEC agar that are used for the selective nature, and for the color change such as what makes the colonies Mauve color?

    1. Hello Amete, sorry for replying to you late. Did you find the answer for your query? We are not using chromogenic agar in our diagnostic laboratory so far, we have managed to purchase some for teaching purposes only.

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