Skip to main content
NCBI home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1997 Sep;35(9):2266–2269. doi: 10.1128/jcm.35.9.2266-2269.1997

Comparative evaluation of modified trichrome and Uvitex 2B stains for detection of low numbers of microsporidial spores in stool specimens.

R Ignatius 1, S Henschel 1, O Liesenfeld 1, U Mansmann 1, W Schmidt 1, S Köppe 1, T Schneider 1, W Heise 1, U Futh 1, E O Riecken 1, H Hahn 1, R Ullrich 1
PMCID: PMC229951  PMID: 9276399

Abstract

At present, the laboratory diagnosis of intestinal infections caused by microsporidia depends on the detection of the typical spores either with a modified trichrome stain (MTS) or by staining with fluorochromes. The purpose of the present study was (i) to compare staining with MTS (MTS method) and the staining with the fluorochrome Uvitex 2B (U2B method) with respect to their sensitivities and specificities, particularly in the presence of low numbers of spores, and (ii) to evaluate their reliabilities under routine laboratory conditions. First, 30 negative human stool specimens as well as 30 specimens enriched with a low concentration of microsporidial spores were examined. The U2B and MTS methods detected 27 and 30, of the positive samples, respectively (95% confidence intervals for sensitivity, 0.73 to 0.98 for the U2B method and 0.88 to 1.00 for the MTS method) without yielding false-positive results (95% confidence intervals for specificity, 0.88 to 1.00 for the MTS and U2B methods). In addition, analysis of serial dilutions of 17 stool specimens from AIDS patients containing microsporidia revealed comparable detection thresholds (P = 0.52) for both methods. Finally, 40 slides prepared from one stool specimen containing very few microsporidia and 40 negative slides were included in the routine diagnostic program during 1 month in order to monitor laboratory handling and run-to-run variations. Again, both methods exhibited comparable sensitivities (95% confidence intervals, 0.83 to 0.99 for the MTS method and 0.91 to 1.00 for the U2B method) and specificities (95% confidence intervals, 0.91 to 1.00 for the MTS and U2B methods). In conclusion, MTS and U2B methods are equally useful in the diagnosis of microsporidiosis. However, since detection thresholds for both methods differed considerably in all diluted stool specimens, performance of a combination of both methods may be more sensitive than the performance of only one procedure in the event of very low numbers of microsporidial spores.

Full Text

The Full Text of this article is available as a PDF (130.6 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. Baselski V. S., Robison M. K., Pifer L. W., Woods D. R. Rapid detection of Pneumocystis carinii in bronchoalveolar lavage samples by using Cellufluor staining. J Clin Microbiol. 1990 Feb;28(2):393–394. doi: 10.1128/jcm.28.2.393-394.1990. [DOI] [PMC free article] [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. DeGirolami P. C., Ezratty C. R., Desai G., McCullough A., Asmuth D., Wanke C., Federman M. Diagnosis of intestinal microsporidiosis by examination of stool and duodenal aspirate with Weber's modified trichrome and Uvitex 2B strains. J Clin Microbiol. 1995 Apr;33(4):805–810. doi: 10.1128/jcm.33.4.805-810.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. Didier E. S., Didier P. J., Friedberg D. N., Stenson S. M., Orenstein J. M., Yee R. W., Tio F. O., Davis R. M., Vossbrinck C., Millichamp N. Isolation and characterization of a new human microsporidian, Encephalitozoon hellem (n. sp.), from three AIDS patients with keratoconjunctivitis. J Infect Dis. 1991 Mar;163(3):617–621. doi: 10.1093/infdis/163.3.617. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Field A. S., Hing M. C., Milliken S. T., Marriott D. J. Microsporidia in the small intestine of HIV-infected patients. A new diagnostic technique and a new species. Med J Aust. 1993 Mar 15;158(6):390–394. [PubMed] [Google Scholar]
  10. 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]
  11. Ignatius R., Lehmann M., Miksits K., Regnath T., Arvand M., Engelmann E., Futh U., Hahn H., Wagner J. A new acid-fast trichrome stain for simultaneous detection of Cryptosporidium parvum and microsporidial species in stool specimens. J Clin Microbiol. 1997 Feb;35(2):446–449. doi: 10.1128/jcm.35.2.446-449.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. 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]
  14. Luna V. A., Stewart B. K., Bergeron D. L., Clausen C. R., Plorde J. J., Fritsche T. R. Use of the fluorochrome calcofluor white in the screening of stool specimens for spores of microsporidia. Am J Clin Pathol. 1995 May;103(5):656–659. doi: 10.1093/ajcp/103.5.656. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. 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]
  17. 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]
  18. 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]
  19. 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]
  20. Velásquez J. N., Carnevale S., Guarnera E. A., Labbé J. H., Chertcoff A., Cabrera M. G., Rodríguez M. I. Detection of the microsporidian parasite Enterocytozoon bieneusi in specimens from patients with AIDS by PCR. J Clin Microbiol. 1996 Dec;34(12):3230–3232. doi: 10.1128/jcm.34.12.3230-3232.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Vávra J., Dahbiová R., Hollister W. S., Canning E. U. Staining of microsporidian spores by optical brighteners with remarks on the use of brighteners for the diagnosis of AIDS associated human microsporidioses. Folia Parasitol (Praha) 1993;40(4):267–272. [PubMed] [Google Scholar]
  22. 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]
  23. 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]
  24. 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]
  25. 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]
  26. 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]
  27. van den Bergh Weerman M. A., van Gool T., Eeftinck Schattenkerk J. K., Dingemans K. P. Electron microscopy as an essential technique for the identification of parasites in aids patients. Eur J Morphol. 1993 Mar-Jun;31(1-2):107–110. [PubMed] [Google Scholar]

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

RESOURCES