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Journal of Clinical Microbiology, February 2005, p. 610-614, Vol. 43, No. 2
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.2.610-614.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Detection and Identification of Enterocytozoon bieneusi and Encephalitozoon Species in Stool and Urine Specimens by PCR and Differential Hybridization

Daan W. Notermans,1* Ron Peek,1 Menno D. de Jong,1,{dagger} Ellen M. Wentink-Bonnema,1 René Boom,2 and Tom van Gool1,3

Section Parasitology,1 Section Clinical Virology, Department of Medical Microbiology, Academic Medical Center, Amsterdam,2 Department of Parasitology, Harbour Hospital, Rotterdam, The Netherlands3

Received 5 July 2004/ Returned for modification 18 August 2004/ Accepted 22 October 2004

Several species of microsporidia can cause disease in humans in both immunocompromised and immunocompetent individuals. Enterocytozoon bieneusi and Encephalitozoon intestinalis are most commonly associated with chronic diarrhea. All Encephalitozoon species, including E. intestinalis, E. hellem, and E. cuniculi, also cause disseminated infections. As distinctive treatment options are available for the different genera, identification is clinically important. We evaluated a PCR with primers directed to a conserved region of the small subunit rRNA gene of microsporidia. Hybridization with a generic microsporidium probe and specific probes for each of the four different species was used for identification. Probes were labeled with ruthenium and detected by electrochemiluminescence. The sensitivity of the assay was tested with plasmids containing the region of interest from each of the four different species and Vittaforma corneae as a control. In addition, the assay was tested with feces spiked with cultured spores from each of the three Encephalitozoon species and V. corneae. An analytical sensitivity of 3.5 x 102 to 3.5 x 103 spores per g of feces, corresponding to 17 to 170 gene copies per PCR, was found, which is several orders of magnitude more sensitive than microscopy after Uvitex 2B fluorescent staining. Stool samples from 22 microscopically diagnosed patients and from 61 uninfected controls were evaluated, showing a sensitivity of at least 95% and a specificity of 100% compared to microscopy. The method was further tested by spiking urine samples with spores of the different Encephalitozoon species.

* Corresponding author. Mailing address: Department of Medical Microbiology, Academic Medical Center, P.O. Box 22660, 1100 DD Amsterdam, The Netherlands. Phone: 31 20 566 3026. Fax: 31 20 566 9745. E-mail: D.W.Notermans{at}amc.uva.nl.

{dagger} Present address: Oxford University Clinical Research Unit, Hospital for Tropical Diseases, Ho Chi Minh City, Vietnam.

Journal of Clinical Microbiology, February 2005, p. 610-614, Vol. 43, No. 2
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.2.610-614.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Bart, A., Wentink-Bonnema, E. M., Heddema, E. R., Buijs, J., van Gool, T. (2008). Frequent Occurrence of Human-Associated Microsporidia in Fecal Droppings of Urban Pigeons in Amsterdam, The Netherlands. Appl. Environ. Microbiol. 74: 7056-7058 [Abstract] [Full Text]  
  • Peek, R., Delbac, F., Speijer, D., Polonais, V., Greve, S., Wentink-Bonnema, E., Ringrose, J., van Gool, T. (2005). Carbohydrate Moieties of Microsporidian Polar Tube Proteins Are Targeted by Immunoglobulin G in Immunocompetent Individuals. Infect. Immun. 73: 7906-7913 [Abstract] [Full Text]  


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