Microsporidia and Candida spores: their discrimination by Calcofluor, trichrome-blue and methylene-blue combination staining.
Faeces of immunocompromised patients are often contaminated with the chitin-containing spores of microsporidia and Candida, which exclude the use of the chitin-specific fluorescent brightener Calcofluor white M2R for the identification of microsporidian spores. We developed a combination staining of Calcofluor white M2R with modified trichrome-blue staining and subsequent methylene-blue incubation which permits discrimination between these two types of spores. As a basis for diagnosis, a difference in the fluorescence pattern (365-440 nm) is combined with a difference in the light microscopic staining pattern. Under fluorescence conditions microsporidia spores have a spotted, brilliant white Calcofluor fluorescence and can easily be identified, while Candida spores show a reddish purple colour. Under the light microscope microsporidian spores show a light red colour with nonstained vacuole spots or strips in contrast to the yeast spores with their red-brown colour. This combination technique offers a highly specific means for the diagnosis of microsporidia spores in faeces.
Schottelius
J
J
Department of Parasitology, Bernhard-Nocht-Institute for Tropical Medicine, Hamburg, Germany.
Kuhn
E M
EM
Enriquez
R
R
D000818:Animals; D001391:Azo Compounds; D001557:Benzenesulfonates; D002176:Candida albicans / Q000254:growth & development / Q000302:isolation & purification*; D002177:Candidiasis / Q000175:diagnosis; D002522:Chlorocebus aethiops; D004801:Eosine Yellowish-(YS); D005243:Feces / Q000382:microbiology; D005456:Fluorescent Dyes; D006801:Humans; D016867:Immunocompromised Host; D008253:Macaca mulatta; D008739:Methyl Green; D008751:Methylene Blue; D016816:Microsporida / Q000254:growth & development / Q000302:isolation & purification*; D016881:Microsporidiosis / Q000175:diagnosis; D012680:Sensitivity and Specificity; D013170:Spores / Q000302:isolation & purification; D013194:Staining and Labeling / Q000592:standards; D014709:Vero Cells
D001391:Azo Compounds; D001557:Benzenesulfonates; D005456:Fluorescent Dyes; C017768:trichrome stain; C007061:C.I. Fluorescent Brightening Agent 28; D008739:Methyl Green; D008751:Methylene Blue; D004801:Eosine Yellowish-(YS)