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Microsporidia and Candida spores: their discrimination by Calcofluor, trichrome-blue and methylene-blue combination

J Schottelius1, E M Kuhn, R Enriquez

  • 1Department of Parasitology, Bernhard-Nocht-Institute for Tropical Medicine, Hamburg, Germany.

Tropical Medicine & International Health : TM & IH
|August 6, 2000
PubMed

Insights

A new staining method differentiates microsporidia from Candida in patient faeces. This technique combines Calcofluor white M2R fluorescence with trichrome-blue and methylene-blue staining for accurate diagnosis.

Area of Science:

  • Medical Mycology
  • Parasitology
  • Diagnostic Microbiology

Background:

  • Immunocompromised patients are susceptible to opportunistic infections.
  • Microsporidia and Candida species are common opportunistic pathogens found in faeces.
  • Calcofluor white M2R, a chitin-specific stain, is limited by cross-reactivity with Candida spores.

Purpose of the Study:

  • To develop a reliable diagnostic method for microsporidia spores in faeces.
  • To overcome the limitations of Calcofluor white M2R staining in mixed infections.
  • To differentiate microsporidia from Candida spores using a combination staining technique.

Main Methods:

  • Developed a combination staining protocol using Calcofluor white M2R, modified trichrome-blue, and methylene-blue.
  • Utilized differences in fluorescence patterns (365-440 nm) for initial identification.
  • Employed light microscopy staining characteristics for definitive differentiation.

Main Results:

  • Microsporidia spores exhibit brilliant white fluorescence with Calcofluor white M2R, while Candida spores appear reddish-purple.
  • Under light microscopy, microsporidia spores stain light red with unstained vacuole spots, distinct from red-brown Candida spores.
  • The combined method allows clear discrimination between microsporidia and Candida spores.

Conclusions:

  • The developed combination staining technique provides a highly specific method for diagnosing microsporidia in faecal samples.
  • This approach enhances diagnostic accuracy, particularly in immunocompromised patients with potential co-infections.
  • The method offers a valuable tool for clinical laboratories dealing with parasitic and fungal infections.

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