Evaluation of Fungifluor, Calcofluor White, and Fungiqual A fluorochrome stains in biopsy imprints and paraffin biopsy sections revealed all three stains to be rapid and sensitive for the detection of microsporidial spores. The spores are appreciated as small refractile bodies in haematoxylin and eosin stain with an unstained area of spore wall, larger than their actual size in modified Warthin-Starry stain and are Gram positive, although some variability in the intensity of staining can be noted. 3 The various stains evaluated on tissue sections include chromotrope 2R modified trichrome, haematoxylin and eosin, Gram and Giemsa stains on jejunal biopsies. The stains that are used on cytologic preparations may not show similar results in biopsy specimens, due to inherent processing artifacts produced and background staining. Most of the stains described in various reports are based on observations on cytologic preparations. They appear as oval to piriform spores measuring 2–7 μm in length and 1.5 to 5 μm in width.
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Light microscopic examination remains the standard test for the diagnosis of microsporidiosis, and considerable progress has been made in the development of additional stains for the diagnosis of these organisms. It is important for pathologists and microbiologists to be knowledgeable regarding the identification of these agents in biopsy, autopsy and cytology specimens. Examples include cornea, conjunctiva, skeletal muscle, small and large intestine, liver, gallbladder, bile duct, pancreatic duct, omentum, kidney, ureter, bladder, prostate, and trachea or bronchi. In most of cases diagnosis of microspoirdiosis could be made by examination of biopsy and autopsy specimens. Two distinct clinical entities of this disease in the eye have been described: deep corneal stromal infection and superficial keratoconjunctivitis. Microsporidia is an important cause of morbidity, and occasionally, mortality in patients with AIDS. In humans, microsporidia are opportunistic pathogens that cause gastrointestinal, sinus, pulmonary, muscular, renal and ocular diseases. Microsporidia is a nontaxonomic designation used to refer to a group of obligate intracellular protists belonging to the phylum Microspora. ConclusionĬonsidering the ease of performance, cost effectiveness and rapidity of the technique, 1% acid fast stain and Gram's chromotrope stain are ideal for the detection of microsporidia. 1% acid fast, Gram's chromotrope and GMS stains provided a reliable diagnosis of microsporidia as diagnostic waist band could be identified and good contrast helped distinguish the spores from inflammatory debris. ResultsĪll sections showed microsporidial spores as 3 – 5 μm, oval bodies. The stained sections were analyzed for the spore characteristics in terms of size, shape, color contrast, cell wall morphology, waist band in cytoplasm and ease of detection. Further sections were prospectively stained with calcofluor white, Gram, Giemsa, Masson's trichrome, acridine orange, Gomori's methenamine silver, Gram's chromotrope and modified acid fast stain. Methodsįour cases diagnosed with microsporidiosis on Hematoxylin and Eosin and Periodic Acid Schiff's stained sections of the corneal button between January 2002 and December 2004, were included. Hence we wanted to evaluate various stains for their ability to detect microsporidia in corneal tissue sections. There is limited data on comparing stains in the detection of microsporidia in corneal biopsies.