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Clinical and Diagnostic Laboratory Immunology, March 1999, p. 243-246, Vol. 6, No. 2
1071-412X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

A Powerful DNA Extraction Method and PCR for Detection of Microsporidia in Clinical Stool Specimens

Andreas Müller,1 Kerstin Stellermann,1 Pia Hartmann,1 Matthias Schrappe,2 Gerd Fätkenheuer,1 Bernd Salzberger,1 Volker Diehl,1 and Caspar Franzen1,*

Department of Internal Medicine I1 and Quality Management,2 University of Cologne, D-50924 Cologne, Germany

Received 29 June 1998/Returned for modification 7 October 1998/Accepted 23 November 1998

The diagnosis of intestinal microsporidiosis has traditionally depended on direct visualization of the parasite in stool specimens or intestinal biopsy samples by light and/or electron microscopy. Limited information about the specificity and sensitivity of PCR for the detection microsporidia in clinical stool specimens is available. To establish a sensitive and specific method for the detection of microsporidia in clinical samples, we studied clinical stool specimens of 104 randomly selected human immunodeficiency virus-infected patients with diarrhea to compare light microscopy and PCR. Fluorochrome Uvitex 2B staining was used for light microscopy. To raise the sensitivity of PCR, we used a powerful and fast DNA extraction method including stool sedimentation, glass bead disruption, and proteinase K and chitinase digestion. PCR was performed with primer pairs V1-PMP2, V1-EB450, and V1-SI500, and the nature of the PCR products was confirmed by Southern blot hybridization. Microsporidiosis was diagnosed by light microscopy in eight patients. Ten patients tested positive for microsporidiosis by PCR. Enterocytozoon bieneusi was found in seven cases, and Encephalitozoon intestinalis was found in four cases. In one case a double infection with E. bieneusi and E. intestinalis was diagnosed by PCR, whereas light microscopy showed only E. bieneusi infection. PCR testing of stool specimens is useful for diagnosis and species differentiation of intestinal microsporidiosis in HIV patients.

* Corresponding author. Mailing address: Department of Internal Medicine I, University of Cologne, Joseph-Stelzmann Str. 9, D-50924 Cologne, Germany. Phone: 49-221-478-4433. Fax: 49-221-478-6456. E-mail: Caspar.Franzen{at}Uni-Koeln.de.

Clinical and Diagnostic Laboratory Immunology, March 1999, p. 243-246, Vol. 6, No. 2
1071-412X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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