PMID: 9435086Jan 22, 1998Paper

PCR amplification and species determination of microsporidia in formalin-fixed feces after immunomagnetic separation

Applied and Environmental Microbiology
S E DowdI L Pepper

Abstract

The term microsporidia is used to describe several species of opportunistic protozoan parasites. Encephalitozoon intestinalis and Enterocytozoon bieneusi have been found in stools of more than 40% of AIDS patients with diarrhea. Diagnosis of infection with these small protozoans has been difficult, and until recently their occurrence has not been well documented. Formalin is widely used to preserve clinical specimens, but due to the nature of the fixation process, subsequent analysis, especially analysis by the PCR, is difficult. This study evaluated methods used to prepare formalin-fixed fecal specimens for PCR amplification of microsporidial DNA. Two methods were devised to allow PCR detection and subsequent identification of microsporidia in formalin-fixed fecal specimens to the species level. One method involved immunomagnetic separation to concentrate microsporidial spores from fecal specimens. In the second method Chelex resin (Bio-Rad, Hercules, Calif.) was used to remove inhibitory substances, followed by a DNA concentration step. Both methods resulted in reproducible, confirmed detection of microsporidia in formalinized fecal specimens and subsequent species determination by PCR sequencing. The detection sensitivity wa...Continue Reading

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Citations

Jan 26, 2010·Biochemical Genetics·Arizumi KikuchiShinichi Itoh
Oct 14, 2009·Clinical Microbiology Reviews·D StarkJ T Ellis
Jun 2, 2011·PLoS Neglected Tropical Diseases·Bohumil SakKamila Saková
Jun 16, 2005·Expert Review of Anti-infective Therapy·Elizabeth S DidierPeter J Didier
May 9, 2000·Clinical and Diagnostic Laboratory Immunology·S CarnevaleM I Rodríguez
Feb 9, 2000·Journal of Applied Microbiology·Y HuJ C Meitzler
Dec 20, 2014·Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc·Bailey L WilbertsEric R Burrough

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