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Journal of Clinical Microbiology, July 1999, p. 2317-2322, Vol. 37, No. 7
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Detection by an Immunofluorescence Test of Encephalitozoon intestinalis Spores in Routinely Formalin-Fixed Stool Samples Stored at Room Temperature

H. Moura,1,2 F. C. Sodre,1,3 F. J. Bornay-Llinares,4 G. J. Leitch,5 T. Navin,1 S. Wahlquist,1 R. Bryan,1 I. Meseguer,4 and G. S. Visvesvara1,*

Division of Parasitic Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Public Health Service, Department of Health and Human Services,1 and Department of Physiology, Morehouse School of Medicine,5 Atlanta, Georgia; DPL, Faculdade de Ciências Médicas, Universidade do Estado do Rio de Janeiro, and Hospital Evandro Chagas, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro,2 and Departamento de Patologia, Faculdade de Medicina, Universidade Federal Fluminense, Niteroi,3 Brazil; and Unidad de Biotecnología Microbiana, Centro de Bioinginiería, Universidad Miguel Hernández, Elche, Alicante, Spain4

Received 12 February 1999/Returned for modification 23 March 1999/Accepted 12 April 1999

Of the several microsporidia that infect humans, Enterocytozoon bieneusi is known to cause a gastrointestinal disease whereas Encephalitozoon intestinalis causes both a disseminated and an intestinal disease. Although several different staining techniques, including the chromotrope technique and its modifications, Uvitex 2B, and the quick-hot Gram-chromotrope procedure, detect microsporidian spores in fecal smears and other clinical samples, they do not identify the species of microsporidia. A need for an easily performed test therefore exists. We reevaluated 120 stool samples that had been found positive for microsporidia previously, using the quick-hot Gram-chromotrope technique, and segregated them into two groups on the basis of spore size. We also screened the smears by immunofluorescence microscopy, using a polyclonal rabbit anti-E. intestinalis serum at a dilution of 1:400. Spores in 29 (24.1%) of the 120 samples fluoresced brightly, indicating that they were E. intestinalis spores. No intense background or cross-reactivity with bacteria, yeasts, or other structures in the stool samples was seen. Additionally, the numbers of spores that fluoresced in seven of these samples were substantially smaller than the numbers of spores that were present in the stained smears, indicating that these samples were probably derived from patients with mixed infections of Enterocytozoon bieneusi and E. intestinalis. Because a 1:400 dilution of this serum does not react with culture-grown Encephalitozoon hellem, Encephalitozoon cuniculi, or Vittaforma corneae or with Enterocytozoon bieneusi spores in feces, we concluded that an immunofluorescence test using this serum is a good alternative for the specific identification of E. intestinalis infections.

* Corresponding author. Mailing address: Division of Parasitic Diseases, M.S.-F-13, Centers for Disease Control and Prevention, 4770 Buford Hwy. NE, Atlanta, GA 30341-3724. Phone: (770) 488-4417. Fax: (770) 488-4253. E-mail: GSV1{at}CDC.GOV.

Journal of Clinical Microbiology, July 1999, p. 2317-2322, Vol. 37, No. 7
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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