Skip to main content
NCBI home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1995 Apr;33(4):805–810. doi: 10.1128/jcm.33.4.805-810.1995

Diagnosis of intestinal microsporidiosis by examination of stool and duodenal aspirate with Weber's modified trichrome and Uvitex 2B strains.

P C DeGirolami 1, C R Ezratty 1, G Desai 1, A McCullough 1, D Asmuth 1, C Wanke 1, M Federman 1
PMCID: PMC228045  PMID: 7540626

Abstract

Severe, chronic diarrhea is a frequent complication of human immunodeficiency virus disease, and intestinal microsporidiosis is being recognized with increasing frequency in patients with AIDS. Noninvasive, cost-effective techniques are needed to optimize its diagnosis. Weber's modified trichrome stain (MTS) and the fluorochrome Uvitex 2B stain were used to detect microsporidial spores in smears of stool and duodenal aspirate (DA) samples received from human immunodeficiency virus-infected patients for examination for ova and parasites. Of 305 samples (292 stool and 13 DA samples) from 140 patients examined by MTS, 83 samples from 26 (18.6%) of the patients were positive for microsporidia (23 patients diagnosed initially and 3 diagnosed upon review). A subset of the samples studied by MTS consisting of 108 smears of stool and DA specimens from 60 patients was examined by Uvitex 2B. All 44 samples positive by MTS were also positive by Uvitex 2B. In addition, seven specimens and three patients were initially detected as positive by Uvitex 2B only (all three patients were positive also by MTS upon review). Confirmation of the diagnosis was obtained for 24 of 26 smear-positive patients by duodenal biopsy and/or stool transmission electron microscopy. Of 114 patients with stained smears negative for microsporidia, 23 had duodenal biopsies which showed no microsporidia. For the 43 patients who underwent duodenal biopsy, the sensitivity of both the MTS and the Uvitex 2B methods compared with biopsy results was 100%. Of six patients with negative duodenal biopsies and positive stained smears, four had microsporidia demonstrated by stool transmission electron microscopy.(ABSTRACT TRUNCATED AT 250 WORDS)

Full Text

The Full Text of this article is available as a PDF (843.3 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aldras A. M., Orenstein J. M., Kotler D. P., Shadduck J. A., Didier E. S. Detection of microsporidia by indirect immunofluorescence antibody test using polyclonal and monoclonal antibodies. J Clin Microbiol. 1994 Mar;32(3):608–612. doi: 10.1128/jcm.32.3.608-612.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Asmuth D. M., DeGirolami P. C., Federman M., Ezratty C. R., Pleskow D. K., Desai G., Wanke C. A. Clinical features of microsporidiosis in patients with AIDS. Clin Infect Dis. 1994 May;18(5):819–825. doi: 10.1093/clinids/18.5.819. [DOI] [PubMed] [Google Scholar]
  3. Bryan R. T., Cali A., Owen R. L., Spencer H. C. Microsporidia: opportunistic pathogens in patients with AIDS. Prog Clin Parasitol. 1991;2:1–26. [PubMed] [Google Scholar]
  4. Cali A., Kotler D. P., Orenstein J. M. Septata intestinalis N. G., N. Sp., an intestinal microsporidian associated with chronic diarrhea and dissemination in AIDS patients. J Eukaryot Microbiol. 1993 Jan-Feb;40(1):101–112. doi: 10.1111/j.1550-7408.1993.tb04889.x. [DOI] [PubMed] [Google Scholar]
  5. Cali A., Owen R. L. Intracellular development of Enterocytozoon, a unique microsporidian found in the intestine of AIDS patients. J Protozool. 1990 Mar-Apr;37(2):145–155. doi: 10.1111/j.1550-7408.1990.tb05885.x. [DOI] [PubMed] [Google Scholar]
  6. Canning E. U., Hollister W. S. Enterocytozoon bieneusi (Microspora): prevalence and pathogenicity in AIDS patients. Trans R Soc Trop Med Hyg. 1990 Mar-Apr;84(2):181–186. doi: 10.1016/0035-9203(90)90247-c. [DOI] [PubMed] [Google Scholar]
  7. Colebunders R., Lusakumuni K., Nelson A. M., Gigase P., Lebughe I., van Marck E., Kapita B., Francis H., Salaun J. J., Quinn T. C. Persistent diarrhoea in Zairian AIDS patients: an endoscopic and histological study. Gut. 1988 Dec;29(12):1687–1691. doi: 10.1136/gut.29.12.1687. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Connolly G. M., Ellis D. S., Williams J. E., Tovey G., Gazzard B. G. Use of electron microscopy in examination of faeces and rectal and jejunal biopsy specimens. J Clin Pathol. 1991 Apr;44(4):313–316. doi: 10.1136/jcp.44.4.313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Desportes I., Le Charpentier Y., Galian A., Bernard F., Cochand-Priollet B., Lavergne A., Ravisse P., Modigliani R. Occurrence of a new microsporidan: Enterocytozoon bieneusi n.g., n. sp., in the enterocytes of a human patient with AIDS. J Protozool. 1985 May;32(2):250–254. doi: 10.1111/j.1550-7408.1985.tb03046.x. [DOI] [PubMed] [Google Scholar]
  10. Dobbins W. O., 3rd, Weinstein W. M. Electron microscopy of the intestine and rectum in acquired immunodeficiency syndrome. Gastroenterology. 1985 Mar;88(3):738–749. doi: 10.1016/0016-5085(85)90145-3. [DOI] [PubMed] [Google Scholar]
  11. Eeftinck Schattenkerk J. K., van Gool T., van Ketel R. J., Bartelsman J. F., Kuiken C. L., Terpstra W. J., Reiss P. Clinical significance of small-intestinal microsporidiosis in HIV-1-infected individuals. Lancet. 1991 Apr 13;337(8746):895–898. doi: 10.1016/0140-6736(91)90215-b. [DOI] [PubMed] [Google Scholar]
  12. Garcia L. S., Shimizu R. Y., Bruckner D. A. Detection of microsporidial spores in fecal specimens from patients diagnosed with cryptosporidiosis. J Clin Microbiol. 1994 Jul;32(7):1739–1741. doi: 10.1128/jcm.32.7.1739-1741.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kokoskin E., Gyorkos T. W., Camus A., Cedilotte L., Purtill T., Ward B. Modified technique for efficient detection of microsporidia. J Clin Microbiol. 1994 Apr;32(4):1074–1075. doi: 10.1128/jcm.32.4.1074-1075.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kotler D. P., Gaetz H. P., Lange M., Klein E. B., Holt P. R. Enteropathy associated with the acquired immunodeficiency syndrome. Ann Intern Med. 1984 Oct;101(4):421–428. doi: 10.7326/0003-4819-101-4-421. [DOI] [PubMed] [Google Scholar]
  15. Ledford D. K., Overman M. D., Gonzalvo A., Cali A., Mester S. W., Lockey R. F. Microsporidiosis myositis in a patient with the acquired immunodeficiency syndrome. Ann Intern Med. 1985 May;102(5):628–630. doi: 10.7326/0003-4819-102-5-628. [DOI] [PubMed] [Google Scholar]
  16. Lucas S. B., Papadaki L., Conlon C., Sewankambo N., Goodgame R., Serwadda D. Diagnosis of intestinal microsporidiosis in patients with AIDS. J Clin Pathol. 1989 Aug;42(8):885–887. doi: 10.1136/jcp.42.8.885-b. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Modigliani R., Bories C., Le Charpentier Y., Salmeron M., Messing B., Galian A., Rambaud J. C., Lavergne A., Cochand-Priollet B., Desportes I. Diarrhoea and malabsorption in acquired immune deficiency syndrome: a study of four cases with special emphasis on opportunistic protozoan infestations. Gut. 1985 Feb;26(2):179–187. doi: 10.1136/gut.26.2.179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Molina J. M., Sarfati C., Beauvais B., Lémann M., Lesourd A., Ferchal F., Casin I., Lagrange P., Modigliani R., Derouin F. Intestinal microsporidiosis in human immunodeficiency virus-infected patients with chronic unexplained diarrhea: prevalence and clinical and biologic features. J Infect Dis. 1993 Jan;167(1):217–221. doi: 10.1093/infdis/167.1.217. [DOI] [PubMed] [Google Scholar]
  19. Orenstein J. M., Chiang J., Steinberg W., Smith P. D., Rotterdam H., Kotler D. P. Intestinal microsporidiosis as a cause of diarrhea in human immunodeficiency virus-infected patients: a report of 20 cases. Hum Pathol. 1990 May;21(5):475–481. doi: 10.1016/0046-8177(90)90003-n. [DOI] [PubMed] [Google Scholar]
  20. Orenstein J. M., Dieterich D. T., Kotler D. P. Systemic dissemination by a newly recognized intestinal microsporidia species in AIDS. AIDS. 1992 Oct;6(10):1143–1150. doi: 10.1097/00002030-199210000-00013. [DOI] [PubMed] [Google Scholar]
  21. Orenstein J. M. Microsporidiosis in the acquired immunodeficiency syndrome. J Parasitol. 1991 Dec;77(6):843–864. [PubMed] [Google Scholar]
  22. Orenstein J. M., Tenner M., Cali A., Kotler D. P. A microsporidian previously undescribed in humans, infecting enterocytes and macrophages, and associated with diarrhea in an acquired immunodeficiency syndrome patient. Hum Pathol. 1992 Jul;23(7):722–728. doi: 10.1016/0046-8177(92)90339-5. [DOI] [PubMed] [Google Scholar]
  23. Rabeneck L., Gyorkey F., Genta R. M., Gyorkey P., Foote L. W., Risser J. M. The role of Microsporidia in the pathogenesis of HIV-related chronic diarrhea. Ann Intern Med. 1993 Nov 1;119(9):895–899. doi: 10.7326/0003-4819-119-9-199311010-00005. [DOI] [PubMed] [Google Scholar]
  24. Rijpstra A. C., Canning E. U., Van Ketel R. J., Eeftinck Schattenkerk J. K., Laarman J. J. Use of light microscopy to diagnose small-intestinal microsporidiosis in patients with AIDS. J Infect Dis. 1988 Apr;157(4):827–831. doi: 10.1093/infdis/157.4.827. [DOI] [PubMed] [Google Scholar]
  25. Ryan N. J., Sutherland G., Coughlan K., Globan M., Doultree J., Marshall J., Baird R. W., Pedersen J., Dwyer B. A new trichrome-blue stain for detection of microsporidial species in urine, stool, and nasopharyngeal specimens. J Clin Microbiol. 1993 Dec;31(12):3264–3269. doi: 10.1128/jcm.31.12.3264-3269.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Shadduck J. A., Greeley E. Microsporidia and human infections. Clin Microbiol Rev. 1989 Apr;2(2):158–165. doi: 10.1128/cmr.2.2.158. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Shadduck J. A. Human microsporidiosis and AIDS. Rev Infect Dis. 1989 Mar-Apr;11(2):203–207. doi: 10.1093/clinids/11.2.203. [DOI] [PubMed] [Google Scholar]
  28. Wachsmuth E. D. A comparison of the highly selective fluorescence staining of fungi in tissue sections with Uvitex 2B and Calcofluor White M2R. Histochem J. 1988 Apr;20(4):215–221. doi: 10.1007/BF01747466. [DOI] [PubMed] [Google Scholar]
  29. Weber R., Bryan R. T. Microsporidial infections in immunodeficient and immunocompetent patients. Clin Infect Dis. 1994 Sep;19(3):517–521. doi: 10.1093/clinids/19.3.517. [DOI] [PubMed] [Google Scholar]
  30. Weber R., Bryan R. T., Owen R. L., Wilcox C. M., Gorelkin L., Visvesvara G. S. Improved light-microscopical detection of microsporidia spores in stool and duodenal aspirates. The Enteric Opportunistic Infections Working Group. N Engl J Med. 1992 Jan 16;326(3):161–166. doi: 10.1056/NEJM199201163260304. [DOI] [PubMed] [Google Scholar]
  31. Zierdt C. H., Gill V. J., Zierdt W. S. Detection of microsporidian spores in clinical samples by indirect fluorescent-antibody assay using whole-cell antisera to Encephalitozoon cuniculi and Encephalitozoon hellem. J Clin Microbiol. 1993 Nov;31(11):3071–3074. doi: 10.1128/jcm.31.11.3071-3074.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. van Gool T., Hollister W. S., Schattenkerk W. E., Van den Bergh Weerman M. A., Terpstra W. J., van Ketel R. J., Reiss P., Canning E. U. Diagnosis of Enterocytozoon bieneusi microsporidiosis in AIDS patients by recovery of spores from faeces. Lancet. 1990 Sep 15;336(8716):697–698. doi: 10.1016/0140-6736(90)92198-q. [DOI] [PubMed] [Google Scholar]
  33. van Gool T., Snijders F., Reiss P., Eeftinck Schattenkerk J. K., van den Bergh Weerman M. A., Bartelsman J. F., Bruins J. J., Canning E. U., Dankert J. Diagnosis of intestinal and disseminated microsporidial infections in patients with HIV by a new rapid fluorescence technique. J Clin Pathol. 1993 Aug;46(8):694–699. doi: 10.1136/jcp.46.8.694. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES