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A powerful DNA extraction method and PCR for detection of microsporidia in clinical stool specimens.
Clin Diagn Lab Immunol. 1999 Mar; 6(2):243-6.CD

Abstract

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.

Authors+Show Affiliations

Department of Internal Medicine I, University of Cologne, D-50924 Cologne, Germany.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

10066661

Citation

Müller, A, et al. "A Powerful DNA Extraction Method and PCR for Detection of Microsporidia in Clinical Stool Specimens." Clinical and Diagnostic Laboratory Immunology, vol. 6, no. 2, 1999, pp. 243-6.
Müller A, Stellermann K, Hartmann P, et al. A powerful DNA extraction method and PCR for detection of microsporidia in clinical stool specimens. Clin Diagn Lab Immunol. 1999;6(2):243-6.
Müller, A., Stellermann, K., Hartmann, P., Schrappe, M., Fätkenheuer, G., Salzberger, B., Diehl, V., & Franzen, C. (1999). A powerful DNA extraction method and PCR for detection of microsporidia in clinical stool specimens. Clinical and Diagnostic Laboratory Immunology, 6(2), 243-6.
Müller A, et al. A Powerful DNA Extraction Method and PCR for Detection of Microsporidia in Clinical Stool Specimens. Clin Diagn Lab Immunol. 1999;6(2):243-6. PubMed PMID: 10066661.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A powerful DNA extraction method and PCR for detection of microsporidia in clinical stool specimens. AU - Müller,A, AU - Stellermann,K, AU - Hartmann,P, AU - Schrappe,M, AU - Fätkenheuer,G, AU - Salzberger,B, AU - Diehl,V, AU - Franzen,C, PY - 1999/3/6/pubmed PY - 1999/3/6/medline PY - 1999/3/6/entrez SP - 243 EP - 6 JF - Clinical and diagnostic laboratory immunology JO - Clin Diagn Lab Immunol VL - 6 IS - 2 N2 - 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. SN - 1071-412X UR - https://www.unboundmedicine.com/medline/citation/10066661/A_powerful_DNA_extraction_method_and_PCR_for_detection_of_microsporidia_in_clinical_stool_specimens_ L2 - http://cvi.asm.org/cgi/pmidlookup?view=long&pmid=10066661 DB - PRIME DP - Unbound Medicine ER -