Because microsporidia have been increasingly recognized as pathogens of both immunosuppressed and immunocompetent persons, rapid and specific diagnosis of microsporidial infection is needed. Definitive diagnosis of microsporidial infection relies on observing microsporidia in biopsy tissue, bodily fluid specimens, or stool by transmission electron microscopy. This method lacks sensitivity when performed on stool and is laborious and specialized. Detection of microsporidia by light microscopic examination of stained stool specimens from patients with gastrointestinal infection allows rapid diagnosis in a routine clinical microbiology laboratory. Because microsporidia are not detected in traditional ova and parasite examinations of stool specimens, modifications of the trichrome stain are now available. The currently accepted standard protocol for modified trichrome staining of microsporidia includes a 1-hour incubation in the modified trichrome stain. We hope to demonstrate that a rapid staining method, which includes shorter exposure to the modified trichrome stain, will provide comparable or superior results. Different modifications of the trichrome stain and different times and temperatures of incubation will be examined. A device that allows stain to be maintained at 50 degrees to 60 degrees C will be used.
