Mycology – Fungal Culture – Superficial/Dermatophytes

Consistent with O. Reg. 671/92 of the French Language Services Act, laboratory testing information on this page is only available in English because it is scientific or technical in nature and is for use only by qualified health care providers and not by members of the public.

Background

This page provides superficial culture testing information for dermatophytes and other organisms causing superficial infections at Public Health Ontario (PHO). Dermatophytes are the causative agents of ringworm (tinea), onychomycosis and dermatophytosis. As well, there are many non-dermatophyte organisms that may cause superficial infections such as erythrasma and pityriosis versicolor.

This page is for information specific to superficial fungal culture. For information regarding other testing options, refer to the following PHO webpages:

Testing Indications

Suspicion of superficial fungal infection, including those that have not been resolved with topical treatments.

Acceptance/Rejection Criteria

Swabs are not a suitable method to collect specimens for superficial fungal culture and will be rejected.

Requests for susceptibility testing of dermatophytes is not a routine test and requires prior approval with the PHO Microbiologist.  Call PHO Customer Service Centre for more information 1-877-604-4567.

Specimen Collection and Handling

Ensure that the patient has not used anti-fungal medications for the previous three days before specimen collection.

Specimen Requirements

Test Requested Required Requisition(s) Specimen Type Minimum Volume Collection Kit

Superficial microscopy and fungal culture (for dermatophytes)

- Skin scrapings

- Nail scrapings /clippings

- Hair (root ends)

Visible on the black paper collection kit
(see submission and collection note # 3 below)

Fungus Culture Kit (Order # 390048)

Rule out erythrasma

Skin scrapings from intertriginous areas (i.e. axilla, anogenital regions, skin folds of the breasts, between toe or finger digits)

Visible on the black paper collection kit
(see submission and collection note # 3 below)

Fungus Culture Kit (Order # 390048)

Rule out tinea versicolor

- Hair (root ends)

- Skin scraping from body trunk (i.e. back , chest), neck, arms, face

Visible on the black paper collection kit
(see submission and collection note # 3 below)

Fungus Culture Kit (Order # 390048)

Submission and Collection Notes

1

Complete all fields of the requisition form, fields a-d are mandatory.

  1. Test(s) requests and indications for testing
  2. Specimen source (with details e.g., skin scraping from which part of the body)
  3. Collection date and time
  4. Immune status – VERY important to note on the requisition if patient has had a transplant, has a haematological malignancy, or is otherwise immunocompromised (including burn wounds, diabetes etc.) as this will impact work up and reporting practices
  5. Current antifungal therapy and/or clinical diagnosis.
  6. Requests for specific organisms of concern (Trichophyton indotineae, erythrasma, tinea versicolor etc.)
  7. Any indications of previous treatment and/or treatment failure.
2

For clinical specimens, label the specimen container(s) with the patient’s first and last name, date of collection, and one other unique identifier such as the patient’s date of birth or Health Card Number. For additional information see: Criteria for Acceptance of Patient Specimens. Failure to provide this information may result in test request rejection or testing delay.

3

Quantity of specimen is crucial – the greater the amount of specimen, the higher the possibility  of isolating the causative agent.

Limitations

  • Specimen volume must be adequate to perform both direct microscopy and culture. Refer to special instructions below on how low volume or low quantity specimens are handled.
  • If specimen does not have specified source and type, appropriate laboratory examination technique may not be applied.

Storage and Transport

Specimens should be stored at room temperature following collection and shipped to Public Health Ontario as soon as possible. All clinical specimens must be shipped in accordance to the Transportation of Dangerous Good Act.

Special Instructions

Specifically note on the requisition if rule-out erythrasma or tinea versicolor (TV) are requested. If sufficient specimen is received, fungal culture will also be performed to detect agents of superficial mycoses and to rule out possible double infections.

Specimens received for rule-out erythrasma or TV that are deemed to be of low volume or quantity will have microscopy prioritized as these organisms are detectable via microscopic examination only.

Specimens received for superficial fungal culture that are deemed to be of low volume or quantity will have culture prioritized.

Requisitions and Kit Ordering

Test Frequency and Turnaround Time (TAT)

Superficial fungal culture is performed Monday to Friday at PHO’s laboratory- Toronto, Orillia, Thunder Bay and Timmins sites

The erythrasma test is performed  daily Monday to Friday at PHO’s laboratory –Toronto only.

Turnaround Time for microscopy: 4 business days from receipt at PHO’s laboratory sites.

Turnaround Time for culture report: minimum of 10 business days (2 weeks) from receipt at PHO’s laboratory sites.

Turnaround Time for Identification of isolates grown in culture: dependent on the organism(s)

Test Methods

Direct microscopic examination of specimens received for ruling-out superficial fungi and or tinea versicolor is performed using calcofluor-white and potassium hydroxide, while specimens received for ruling-out erythrasma is performed in addition using modified methylene blue stain.

Erythrasma is not a fungal infection and is caused by a bacterium called Corynebacterium minutissimum.  Diagnosis is typically completed in the Mycology Laboratory.

Identification of yeasts and filamentous fungi isolated in culture uses various conventional phenotypic methods, including microscopic morphologic examination, interpretation of biochemical reactions and API strip reactions,  growth temperature and culture morphologic characteristics. Molecular methods such as sequencing and matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) are used when phenotypic methods shows result discrepancy.

Algorithm

If sufficient quantity of specimen is available, both direct microscopy and culture are performed simultaneously.  If quantity is not sufficient, priorities are determined based on what is requested specifically to be ruled-out. Refer to the special instruction above on how examinations are prioritized when a specimen with low quantity is received.

Yeast identification is not routine unless seen in the direct microscopic examination or for patients who are immunocompromised.

Interpretation

The following table provides possible test results with associated interpretations:

Microscopic Result

Interpretation

Final Report Comment

Characteristic of Corynebacterium minutissimum, causative agent of erythrasma

Erythrasma confirmed.

Final report based on direct microscopy. 

Culture will be performed and clients will only be notified if a fungus of significance is isolated.

Suggestive of Corynebacterium minutissimum, causative agent of Erythrasma

Possible erythrasma

Final report based on direct microscopy. 

 

Culture will be performed and clients will only be notified if a fungus of significance is isolated.

Negative for erythrasma

Erythrasma ruled-out.

Final report based on direct microscopy. 

Culture will be performed and clients will only be notified if a fungus of significance is isolated.

Characteristic of Malassezia furfur, causative agent of pityriasis versicolor (tinea versicolor, TV)

Pityriasis versicolor (tinea versicolor, TV) confirmed.

Final report based on direct microscopy. 

Culture will be performed and clients will only be notified if a fungus of significance is isolated.

Suggestive of Malassezia furfur, causative agent of pityriasis versicolor (tinea versicolor, TV)

Possible pityriasis versicolor (tinea versicolor, TV)

Final report based on direct microscopy. 

Culture will be performed and clients will only be notified if a fungus of significance is isolated.

Negative for pityriasis versicolor (tinea versicolor, TV)

Possible pityriasis versicolor (tinea versicolor, TV)ruled-out.

Culture will be performed and clients will only be notified if a fungus of significance is isolated.

Mycelium seen

Fungal spores and mycelium  seen

Suggestive of dermatophyte present

Identification of isolate(s) grown in culture to genus/species.

Fungal Filaments seen

Irregular mycelium seen

Yeast cells and filaments seen

Budding yeast cells seen

Presence of yeast or non-dermatophytic fungi in specimen.

Clinical significance is unknown.

Dermatophytes will be ruled-out or identified to genus/species if isolated.

Non-dermatophyte organisms that are known to cause onychomycosis will be identified to genus/species.

Clinically insignificant growth will be reported as ‘non dermatophyte isolated’ or ‘mixture of non dermatophytes isolated’.

Fungal cultures are identified to the genus and species level when applicable.

Reporting

Results are reported to the physician, authorized health care provider (General O. Reg 45/22, s.18) or submitter as indicated on the requisition.

For isolates known to cause reportable diseases: specimens are reported to the Medical Officer of Health as per the Ontario Health Protection and Promotion Act.

References

  1. Clinical and Laboratory Standards Institute (CLSI). Principles and Procedures for Detection and Culture of Fungi in Clinical Specimens. 2nd ed. CLSI guideline M54 (ISBN 978-1-68440-098-0 {Print}; ISBN 978-1-68440-099-7 {Electronic}). Clinical and Laboratory Standards Institute; 2021
  2. Hazen, K., Howell, S.A.. Mycology and antifungal susceptibility testing. In: Leber, A.M. Editor. Clinical Microbiology Procedures Handbook. 4th ed. Washington, D.C.; ASM Press; 2016.
  3. de Hoog, G.S., Guarro, J., Gene, J., Ahmed, S.A., Al-Hatmi, A.M.S., Figueras, M.J., Vitale, R.G.. Atlas of Clinical Fungi. 4th ed. Hilversum; Foundation Atlas of Clinical Fungi; 2020.
  4. Kane, J., Summerbell, R., Sigler, L., Krajden, S., Land, G.. Laboatory Handbook of Dermatophytes: A Clinical Guide and Laboratory Manual of Dermatophytes and Other Filamentous Fungi from Skin, Hair and Nails. USA: Star Publishing Company; 1997
Updated 12 Feb 2025