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Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1997 Feb;35(2):446–449. doi: 10.1128/jcm.35.2.446-449.1997

A new acid-fast trichrome stain for simultaneous detection of Cryptosporidium parvum and microsporidial species in stool specimens.

R Ignatius 1, M Lehmann 1, K Miksits 1, T Regnath 1, M Arvand 1, E Engelmann 1, U Futh 1, H Hahn 1, J Wagner 1
PMCID: PMC229597  PMID: 9003613

Abstract

The detection in stool specimens of Cryptosporidium parvum and microsporidia, the most frequent parasitic pathogens causing diarrhea in AIDS patients, until now has depended on two different staining methods. However, since double infections occur and minimization of laboratory costs is mandatory, development of a method for simultaneous detection of these parasites appeared desirable. We report on a new, inexpensive, and easy-to-perform staining procedure to demonstrate both acid-fast oocysts of C. parvum and other coccidia, as well as microsporidial spores. This acid-fast trichrome stain yields results comparable to those obtained by the Kinyoun and modified trichrome methods and considerably reduces the time necessary for microscopic examination.

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Selected References

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  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. Current W. L., Garcia L. S. Cryptosporidiosis. Clin Microbiol Rev. 1991 Jul;4(3):325–358. doi: 10.1128/cmr.4.3.325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Garcia L. S., Brewer T. C., Bruckner D. A. Fluorescence detection of Cryptosporidium oocysts in human fecal specimens by using monoclonal antibodies. J Clin Microbiol. 1987 Jan;25(1):119–121. doi: 10.1128/jcm.25.1.119-121.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Garcia L. S., Bruckner D. A., Brewer T. C., Shimizu R. Y. Techniques for the recovery and identification of Cryptosporidium oocysts from stool specimens. J Clin Microbiol. 1983 Jul;18(1):185–190. doi: 10.1128/jcm.18.1.185-190.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Hart A. S., Ridinger M. T., Soundarajan R., Peters C. S., Swiatlo A. L., Kocka F. E. Novel organism associated with chronic diarrhoea in AIDS. Lancet. 1990 Jan 20;335(8682):169–170. doi: 10.1016/0140-6736(90)90042-4. [DOI] [PubMed] [Google Scholar]
  9. Højlyng N., Jensen B. N. Respiratory cryptosporidiosis in HIV-positive patients. Lancet. 1988 Mar 12;1(8585):590–591. doi: 10.1016/s0140-6736(88)91384-0. [DOI] [PubMed] [Google Scholar]
  10. López-Vélez R., Tarazona R., Garcia Camacho A., Gomez-Mampaso E., Guerrero A., Moreira V., Villanueva R. Intestinal and extraintestinal cryptosporidiosis in AIDS patients. Eur J Clin Microbiol Infect Dis. 1995 Aug;14(8):677–681. doi: 10.1007/BF01690873. [DOI] [PubMed] [Google Scholar]
  11. Ng E., Markell E. K., Fleming R. L., Fried M. Demonstration of Isospora belli by acid-fast stain in a patient with acquired immune deficiency syndrome. J Clin Microbiol. 1984 Sep;20(3):384–386. doi: 10.1128/jcm.20.3.384-386.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Scholten T. H., Yang J. Evaluation of unpreserved and preserved stools for the detection and identification of intestinal parasites. Am J Clin Pathol. 1974 Oct;62(4):563–567. doi: 10.1093/ajcp/62.4.563. [DOI] [PubMed] [Google Scholar]
  14. Sterling C. R., Arrowood M. J. Detection of Cryptosporidium sp. infections using a direct immunofluorescent assay. Pediatr Infect Dis. 1986 Jan-Feb;5(1 Suppl):S139–S142. doi: 10.1097/00006454-198601001-00022. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Weber R., Sauer B., Lüthy R., Nadal D. Intestinal coinfection with Enterocytozoon bieneusi and Cryptosporidium in a human immunodeficiency virus-infected child with chronic diarrhea. Clin Infect Dis. 1993 Sep;17(3):480–483. doi: 10.1093/clinids/17.3.480. [DOI] [PubMed] [Google Scholar]
  17. Wurtz R. M., Kocka F. E., Peters C. S., Weldon-Linne C. M., Kuritza A., Yungbluth P. Clinical characteristics of seven cases of diarrhea associated with a novel acid-fast organism in the stool. Clin Infect Dis. 1993 Jan;16(1):136–138. doi: 10.1093/clinids/16.1.136. [DOI] [PubMed] [Google Scholar]
  18. Yang J., Scholten T. A fixative for intestinal parasites permitting the use of concentration and permanent staining procedures. Am J Clin Pathol. 1977 Mar;67(3):300–304. doi: 10.1093/ajcp/67.3.300. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. 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]

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