Skip to main content
Emerging Infectious Diseases logoLink to Emerging Infectious Diseases
. 1996 Jul-Sep;2(3):183–191. doi: 10.3201/eid0203.960304

Application of molecular techniques to the diagnosis of microsporidial infection.

D P Fedorko 1, Y M Hijazi 1
PMCID: PMC2626796  PMID: 8903228

Abstract

Microsporidia are now recognized as important pathogens of AIDS patients; the ability of these parasites to cause disease in immunocompetent persons is still being elucidated. Improved diagnostic tests for microsporidial infection are continually being sought for establishing diagnosis in order to avoid laborious electron microscopy studies that require invasively acquired biopsy specimens. Modified trichrome or chemofluorescent stains are useful for detecting microsporidia in bodily fluids and stool specimens, but they do not allow for speciation of microsporidia. Polymerase chain reaction with specific primers will allow the detection and speciation of microsporidia in biopsy tissue, bodily fluids, and stool specimens.

Full Text

The Full Text of this article is available as a PDF (1.1 MB).

Selected References

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

  1. Baker M. D., Vossbrinck C. R., Didier E. S., Maddox J. V., Shadduck J. A. Small subunit ribosomal DNA phylogeny of various microsporidia with emphasis on AIDS related forms. J Eukaryot Microbiol. 1995 Sep-Oct;42(5):564–570. doi: 10.1111/j.1550-7408.1995.tb05906.x. [DOI] [PubMed] [Google Scholar]
  2. Beckers P. J., Derks G. J., Gool T., Rietveld F. J., Sauerwein R. W. Encephalocytozoon intestinalis-specific monoclonal antibodies for laboratory diagnosis of microsporidiosis. J Clin Microbiol. 1996 Feb;34(2):282–285. doi: 10.1128/jcm.34.2.282-285.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bergquist R., Morfeldt-Månsson L., Pehrson P. O., Petrini B., Wasserman J. Antibody against Encephalitozoon cuniculi in Swedish homosexual men. Scand J Infect Dis. 1984;16(4):389–391. doi: 10.3109/00365548409073966. [DOI] [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. Clarridge J. E., 3rd, Karkhanis S., Rabeneck L., Marino B., Foote L. W. Quantitative light microscopic detection of Enterocytozoon bieneusi in stool specimens: a longitudinal study of human immunodeficiency virus-infected microsporidiosis patients. J Clin Microbiol. 1996 Mar;34(3):520–523. doi: 10.1128/jcm.34.3.520-523.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. De Groote M. A., Visvesvara G., Wilson M. L., Pieniazek N. J., Slemenda S. B., daSilva A. J., Leitch G. J., Bryan R. T., Reves R. Polymerase chain reaction and culture confirmation of disseminated Encephalitozoon cuniculi in a patient with AIDS: successful therapy with albendazole. J Infect Dis. 1995 May;171(5):1375–1378. doi: 10.1093/infdis/171.5.1375. [DOI] [PubMed] [Google Scholar]
  7. Deplazes P., Mathis A., Baumgartner R., Tanner I., Weber R. Immunologic and molecular characteristics of Encephalitozoon-like microsporidia isolated from humans and rabbits indicate that Encephalitozoon cuniculi is a zoonotic parasite. Clin Infect Dis. 1996 Mar;22(3):557–559. doi: 10.1093/clinids/22.3.557. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Didier E. S., Orenstein J. M., Aldras A., Bertucci D., Rogers L. B., Janney F. A. Comparison of three staining methods for detecting microsporidia in fluids. J Clin Microbiol. 1995 Dec;33(12):3138–3145. doi: 10.1128/jcm.33.12.3138-3145.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Didier E. S., Rogers L. B., Brush A. D., Wong S., Traina-Dorge V., Bertucci D. Diagnosis of disseminated microsporidian Encephalitozoon hellem infection by PCR-Southern analysis and successful treatment with albendazole and fumagillin. J Clin Microbiol. 1996 Apr;34(4):947–952. doi: 10.1128/jcm.34.4.947-952.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Didier E. S., Shadduck J. A., Didier P. J., Millichamp N., Vossbrinck C. R. Studies on ocular microsporidia. J Protozool. 1991 Nov-Dec;38(6):635–638. [PubMed] [Google Scholar]
  12. Didier E. S., Vossbrinck C. R., Baker M. D., Rogers L. B., Bertucci D. C., Shadduck J. A. Identification and characterization of three Encephalitozoon cuniculi strains. Parasitology. 1995 Nov;111(Pt 4):411–421. doi: 10.1017/s0031182000065914. [DOI] [PubMed] [Google Scholar]
  13. Fedorko D. P., Nelson N. A., Cartwright C. P. Identification of microsporidia in stool specimens by using PCR and restriction endonucleases. J Clin Microbiol. 1995 Jul;33(7):1739–1741. doi: 10.1128/jcm.33.7.1739-1741.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Field A. S., Marriott D. J., Hing M. C. The Warthin-Starry stain in the diagnosis of small intestinal microsporidiosis in HIV-infected patients. Folia Parasitol (Praha) 1993;40(4):261–266. [PubMed] [Google Scholar]
  15. Franzen C., Müller A., Hegener P., Salzberger B., Hartmann P., Fätkenheuer G., Diehl V., Schrappe M. Detection of microsporidia (Enterocytozoon bieneusi) in intestinal biopsy specimens from human immunodeficiency virus-infected patients by PCR. J Clin Microbiol. 1995 Sep;33(9):2294–2296. doi: 10.1128/jcm.33.9.2294-2296.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hartskeerl R. A., Schuitema A. R., deWachter R. Secondary structure of the small subunit ribosomal RNA sequence of the microsporidium Encephalitozoon cuniculi. Nucleic Acids Res. 1993 Mar 25;21(6):1489–1489. doi: 10.1093/nar/21.6.1489. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hartskeerl R. A., Schuitema A. R., van Gool T., Terpstra W. J. Genetic evidence for the occurrence of extra-intestinal Enterocytozoon bieneusi infections. Nucleic Acids Res. 1993 Aug 25;21(17):4150–4150. doi: 10.1093/nar/21.17.4150. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hartskeerl R. A., Van Gool T., Schuitema A. R., Didier E. S., Terpstra W. J. Genetic and immunological characterization of the microsporidian Septata intestinalis Cali, Kotler and Orenstein, 1993: reclassification to Encephalitozoon intestinalis. Parasitology. 1995 Apr;110(Pt 3):277–285. doi: 10.1017/s0031182000080860. [DOI] [PubMed] [Google Scholar]
  19. He Q., Leitch G. J., Visvesvara G. S., Wallace S. Effects of nifedipine, metronidazole, and nitric oxide donors on spore germination and cell culture infection of the microsporidia Encephalitozoon hellem and Encephalitozoon intestinalis. Antimicrob Agents Chemother. 1996 Jan;40(1):179–185. doi: 10.1128/aac.40.1.179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hollister W. S., Canning E. U. An enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to Encephalitozoon cuniculi and its use in determination of infections in man. Parasitology. 1987 Apr;94(Pt 2):209–219. doi: 10.1017/s0031182000053890. [DOI] [PubMed] [Google Scholar]
  21. Hollister W. S., Canning E. U., Willcox A. Evidence for widespread occurrence of antibodies to Encephalitozoon cuniculi (Microspora) in man provided by ELISA and other serological tests. Parasitology. 1991 Feb;102(Pt 1):33–43. doi: 10.1017/s0031182000060315. [DOI] [PubMed] [Google Scholar]
  22. Kotler D. P., Giang T. T., Garro M. L., Orenstein J. M. Light microscopic diagnosis of microsporidiosis in patients with AIDS. Am J Gastroenterol. 1994 Apr;89(4):540–544. [PubMed] [Google Scholar]
  23. Kotler D. P., Orenstein J. M. Prevalence of intestinal microsporidiosis in HIV-infected individuals referred for gastroenterological evaluation. Am J Gastroenterol. 1994 Nov;89(11):1998–2002. [PubMed] [Google Scholar]
  24. MATSUBAYASHI H., KOIKE T., MIKATA I., TAKEI H., HAGIWARA S. A case of Encephalitozoon-like body infection in man. AMA Arch Pathol. 1959 Feb;67(2):181–187. [PubMed] [Google Scholar]
  25. 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]
  26. Ombrouck C., Romestand B., da Costa J. M., Desportes-Livage I., Datry A., Coste F., Bouix G., Gentilini M. Use of cross-reactive antigens of the microsporidian Glugea atherinae for the possible detection of Enterocytozoon bieneusi by western blot. Am J Trop Med Hyg. 1995 Jan;52(1):89–93. doi: 10.4269/ajtmh.1995.52.89. [DOI] [PubMed] [Google Scholar]
  27. Orenstein J. M. Microsporidiosis in the acquired immunodeficiency syndrome. J Parasitol. 1991 Dec;77(6):843–864. [PubMed] [Google Scholar]
  28. Sandfort J., Hannemann A., Gelderblom H., Stark K., Owen R. L., Ruf B. Enterocytozoon bieneusi infection in an immunocompetent patient who had acute diarrhea and who was not infected with the human immunodeficiency virus. Clin Infect Dis. 1994 Sep;19(3):514–516. doi: 10.1093/clinids/19.3.514. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. Silveira H., Canning E. U. Vittaforma corneae n. comb. for the human microsporidium Nosema corneum Shadduck, Meccoli, Davis & Font, 1990, based on its ultrastructure in the liver of experimentally infected athymic mice. J Eukaryot Microbiol. 1995 Mar-Apr;42(2):158–165. doi: 10.1111/j.1550-7408.1995.tb01557.x. [DOI] [PubMed] [Google Scholar]
  31. Visvesvara G. S., Leitch G. J., da Silva A. J., Croppo G. P., Moura H., Wallace S., Slemenda S. B., Schwartz D. A., Moss D., Bryan R. T. Polyclonal and monoclonal antibody and PCR-amplified small-subunit rRNA identification of a microsporidian, Encephalitozoon hellem, isolated from an AIDS patient with disseminated infection. J Clin Microbiol. 1994 Nov;32(11):2760–2768. doi: 10.1128/jcm.32.11.2760-2768.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Visvesvara G., Leitch G. J., Pieniazek N. J., Da Silva A. J., Wallace S., Slemenda S. B., Weber R., Schwartz D. A., Gorelkin L., Wilcox C. M. Short-term in vitro culture and molecular analysis of the microsporidian, Enterocytozoon bieneusi. J Eukaryot Microbiol. 1995 Sep-Oct;42(5):506–510. doi: 10.1111/j.1550-7408.1995.tb05896.x. [DOI] [PubMed] [Google Scholar]
  33. Vossbrinck C. R., Baker M. D., Didier E. S., Debrunner-Vossbrinck B. A., Shadduck J. A. Ribosomal DNA sequences of Encephalitozoon hellem and Encephalitozoon cuniculi: species identification and phylogenetic construction. J Eukaryot Microbiol. 1993 May-Jun;40(3):354–362. doi: 10.1111/j.1550-7408.1993.tb04928.x. [DOI] [PubMed] [Google Scholar]
  34. Vossbrinck C. R., Maddox J. V., Friedman S., Debrunner-Vossbrinck B. A., Woese C. R. Ribosomal RNA sequence suggests microsporidia are extremely ancient eukaryotes. 1987 Mar 26-Apr 1Nature. 326(6111):411–414. doi: 10.1038/326411a0. [DOI] [PubMed] [Google Scholar]
  35. 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]
  36. Weber R., Bryan R. T., Schwartz D. A., Owen R. L. Human microsporidial infections. Clin Microbiol Rev. 1994 Oct;7(4):426–461. doi: 10.1128/cmr.7.4.426. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Weiss L. M., Zhu X., Cali A., Tanowitz H. B., Wittner M. Utility of microsporidian rRNA in diagnosis and phylogeny: a review. Folia Parasitol (Praha) 1994;41(2):81–90. [PubMed] [Google Scholar]
  38. Zhu X., Wittner M., Tanowitz H. B., Kotler D., Cali A., Weiss L. M. Small subunit rRNA sequence of Enterocytozoon bieneusi and its potential diagnostic role with use of the polymerase chain reaction. J Infect Dis. 1993 Dec;168(6):1570–1575. doi: 10.1093/infdis/168.6.1570. [DOI] [PubMed] [Google Scholar]
  39. da Silva A. J., Schwartz D. A., Visvesvara G. S., de Moura H., Slemenda S. B., Pieniazek N. J. Sensitive PCR diagnosis of Infections by Enterocytozoon bieneusi (microsporidia) using primers based on the region coding for small-subunit rRNA. J Clin Microbiol. 1996 Apr;34(4):986–987. doi: 10.1128/jcm.34.4.986-987.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. van Gool T., Canning E. U., Gilis H., van den Bergh Weerman M. A., Eeftinck Schattenkerk J. K., Dankert J. Septata intestinalis frequently isolated from stool of AIDS patients with a new cultivation method. Parasitology. 1994 Sep;109(Pt 3):281–289. doi: 10.1017/s0031182000078318. [DOI] [PubMed] [Google Scholar]

Articles from Emerging Infectious Diseases are provided here courtesy of Centers for Disease Control and Prevention

RESOURCES