A powerful DNA extraction method and PCR for detection of microsporidia in clinical stool specimens.

Important note: Information in this article was accurate in 1999. The state of the art may have changed since the publication date.

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. Unique Identifier : AIDSLINE MED/99167672
Muller A; Stellermann K; Hartmann P; Schrappe M; Fatkenheuer G; Salzberger B; Diehl V; Franzen C; Department of Internal Medicine I, University of Cologne, D-50924; Cologne, Germany.

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.

Keywords: JOURNAL ARTICLE Animal AIDS-Related Opportunistic Infections/DIAGNOSIS Blotting, Southern DNA Primers DNA, Protozoan/*ANALYSIS/ISOLATION & PURIF Feces/*PARASITOLOGY Human Intestinal Diseases, Parasitic/*DIAGNOSIS Microspora Infections/*DIAGNOSIS Microsporida/GENETICS/*ISOLATION & PURIF Polymerase Chain Reaction/METHODS/STANDARDS Reproducibility of Results Sensitivity and Specificity Support, Non-U.S. Gov't
990630
A9960963

AEGiS is a 501(c)3, not-for-profit, tax-exempt, educational corporation. AEGiS is made possible through unrestricted funding from Boehringer Ingelheim, Bridgestone/Firestone Charitable Trust, Bristol-Myers Squibb Company, Elton John AIDS Foundation, Gill Foundation, the National Library of Medicine, Quest Diagnostics, Roche and Trimeris, and donations from users like you. Always watch for outdated information. This article first appeared in 1999. This material is designed to support, not replace, the relationship that exists between you and your doctor.

AEGiS presents published material, reprinted with permission and neither endorses nor opposes any material. All information contained on this website, including information relating to health conditions, products, and treatments, is for informational purposes only. It is often presented in summary or aggregate form. It is not meant to be a substitute for the advice provided by your own physician or other medical professionals. Always discuss treatment options with a doctor who specializes in treating HIV.

Copyright ©1980, 1999. AEGiS. All materials appearing on AEGiS are protected by copyright as a collective work or compilation under U.S. copyright and other laws and are the property of AEGiS, or the party credited as the provider of the content. .