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Clin Diagn Lab Immunol. 1997 July; 4(4): 405–408.
PMCID: PMC170541

Development and application of genetic probes for detection of Enterocytozoon bieneusi in formalin-fixed stools and in intestinal biopsy specimens from infected patients.

Abstract

The microsporidium Enterocytozoon bieneusi is closely linked to wasting and diarrhea in a high proportion of individuals with AIDS. However, its relative contribution to disease is uncertain because diagnosis until recently depended on procedures involving endoscopy. A sensitive PCR technique which amplifies a fragment of the small-subunit rRNA gene of E. bieneusi from formalin-fixed stool samples was developed. Of 80 formalin-fixed stool samples collected from 74 Zimbabweans and 6 U.S. patients who were human immunodeficiency virus positive, 50% tested positive for E. bieneusi by PCR, whereas 24% tested positive for E. bieneusi by light microscopy of trichrome-stained fecal smears. In addition, we describe an in situ hybridization technique which detected and identified E. bieneusi as the causative agent in all six intestinal biopsy specimens tested. Both the PCR and in situ hybridization procedures are sensitive diagnostic tools which will complement currently available techniques and enable the differentiation of E. bieneusi from other microsporidia to be made.

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Selected References

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  • Bryan RT, Cali A, Owen RL, Spencer HC. Microsporidia: opportunistic pathogens in patients with AIDS. Prog Clin Parasitol. 1991;2:1–26. [PubMed]
  • Cali A, Owen RL. Intracellular development of Enterocytozoon, a unique microsporidian found in the intestine of AIDS patients. J Protozool. 1990 Mar-Apr;37(2):145–155. [PubMed]
  • Conteas CN, Sowerby T, Berlin GW, Dahlan F, Nguyen A, Porschen R, Donovan J, LaRiviere M, Orenstein JM. Fluorescence techniques for diagnosing intestinal microsporidiosis in stool, enteric fluid, and biopsy specimens from acquired immunodeficiency syndrome patients with chronic diarrhea. Arch Pathol Lab Med. 1996 Sep;120(9):847–853. [PubMed]
  • da Silva AJ, Schwartz DA, Visvesvara GS, de Moura H, Slemenda SB, Pieniazek NJ. 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. [PMC free article] [PubMed]
  • 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. [PubMed]
  • Dobbins WO, 3rd, Weinstein WM. Electron microscopy of the intestine and rectum in acquired immunodeficiency syndrome. Gastroenterology. 1985 Mar;88(3):738–749. [PubMed]
  • Fedorko DP, Nelson NA, Cartwright CP. Identification of microsporidia in stool specimens by using PCR and restriction endonucleases. J Clin Microbiol. 1995 Jul;33(7):1739–1741. [PMC free article] [PubMed]
  • 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. [PMC free article] [PubMed]
  • Hollister WS, Canning EU, Weidner E, Field AS, Kench J, Marriott DJ. Development and ultrastructure of Trachipleistophora hominis n.g., n.sp. after in vitro isolation from an AIDS patient and inoculation into athymic mice. Parasitology. 1996 Jan;112(Pt 1):143–154. [PubMed]
  • Ilyinskii PO, Daniel MD, Simon MA, Lackner AA, Desrosiers RC. The role of upstream U3 sequences in the pathogenesis of simian immunodeficiency virus-induced AIDS in rhesus monkeys. J Virol. 1994 Sep;68(9):5933–5944. [PMC free article] [PubMed]
  • Kotler DP, Orenstein JM. Prevalence of intestinal microsporidiosis in HIV-infected individuals referred for gastroenterological evaluation. Am J Gastroenterol. 1994 Nov;89(11):1998–2002. [PubMed]
  • Mansfield KG, Carville A, Shvetz D, MacKey J, Tzipori S, Lackner AA. Identification of an Enterocytozoon bieneusi-like microsporidian parasite in simian-immunodeficiency-virus-inoculated macaques with hepatobiliary disease. Am J Pathol. 1997 Apr;150(4):1395–1405. [PubMed]
  • 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]
  • Modigliani R, Bories C, Le Charpentier Y, Salmeron M, Messing B, Galian A, Rambaud JC, 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. [PMC free article] [PubMed]
  • Orenstein JM, Chiang J, Steinberg W, Smith PD, Rotterdam H, Kotler DP. 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. [PubMed]
  • Orenstein JM, Dieterich DT, Kotler DP. Systemic dissemination by a newly recognized intestinal microsporidia species in AIDS. AIDS. 1992 Oct;6(10):1143–1150. [PubMed]
  • Pol S, Romana CA, Richard S, Amouyal P, Desportes-Livage I, Carnot F, Pays JF, Berthelot P. Microsporidia infection in patients with the human immunodeficiency virus and unexplained cholangitis. N Engl J Med. 1993 Jan 14;328(2):95–99. [PubMed]
  • Sandfort J, Hannemann A, Gelderblom H, Stark K, Owen RL, 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. [PubMed]
  • Tzipori S, Carville A, Widmer G, Kotler D, Mansfield K, Lackner A. Transmission and establishment of a persistent infection of Enterocytozoon bieneusi, derived from a human with AIDS, in simian immunodeficiency virus-infected rhesus monkeys. J Infect Dis. 1997 Apr;175(4):1016–1020. [PubMed]
  • Weber R, Bryan RT, Owen RL, Wilcox CM, Gorelkin L, Visvesvara GS. 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. [PubMed]
  • Weber R, Bryan RT, Schwartz DA, Owen RL. Human microsporidial infections. Clin Microbiol Rev. 1994 Oct;7(4):426–461. [PMC free article] [PubMed]
  • Weiss LM, Zhu X, Cali A, Tanowitz HB, Wittner M. Utility of microsporidian rRNA in diagnosis and phylogeny: a review. Folia Parasitol (Praha) 1994;41(2):81–90. [PubMed]
  • Wittner M, Tanowitz HB, Weiss LM. Parasitic infections in AIDS patients. Cryptosporidiosis, isosporiasis, microsporidiosis, cyclosporiasis. Infect Dis Clin North Am. 1993 Sep;7(3):569–586. [PubMed]
  • Zhu X, Wittner M, Tanowitz HB, Kotler D, Cali A, Weiss LM. 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. [PubMed]

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