Purification of Enterocytozoon bieneusi spores from stool specimens by gradient and cell sorting techniques

Journal of Clinical Microbiology
Zuzana KucerovaG S Visvesvara

Abstract

A three-step method for the purification of Enterocytozoon bieneusi spores from stool specimens was developed. The primary process of purification of the spores from bacterial contaminants involved Percoll gradient centrifugation followed by additional separation using cesium chloride density gradient centrifugation. The cesium chloride-isolated spores were further purified using a flow cytometer with cell sorting capabilities. Sorting was performed without the use of antibodies, fluorochromes, or dyes, leaving the sorted spores in their native state, which appears to be less destructive for spores. When quantified by flow cytometry using tubes with known numbers of highly fluorescent polystyrene beads, the sorted material showed a slight decrease in light scatter characteristics compared with the slightly larger Encephalitozoon species spores. Although the overall recovery of the E. bieneusi spores was low, calcofluor and Gram chromotrope staining, indirect immunofluorescence assay, and transmission electron microscopy revealed that the sorted material was highly purified and contained large numbers of E. bieneusi spores and relatively few bacteria and other debris. The sorted material appeared to be sufficiently pure and coul...Continue Reading

References

Sep 1, 1994·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·J SandfortB Ruf
Oct 1, 1994·Clinical Microbiology Reviews·R WeberR L Owen
May 1, 1994·The Journal of Eukaryotic Microbiology·J M AmigóC P Vivarès
May 1, 1997·Clinical and Diagnostic Laboratory Immunology·J K NicholsonT Denny
Jan 16, 1999·Journal of Clinical Microbiology·D M MossG S Visvesvara
Aug 26, 1999·The Journal of Eukaryotic Microbiology·L C GreenE S Didier
Nov 24, 1999·Journal of Clinical Microbiology·I AccoceberryA Datry
Nov 27, 1999·Journal of Travel Medicine·R López-VélezC del Aguila
May 5, 2000·European Journal of Clinical Microbiology & Infectious Diseases : Official Publication of the European Society of Clinical Microbiology·S MetgeS Bretagne
Jul 13, 2000·Journal of Biochemical and Biophysical Methods·H Pertoft
Oct 12, 2000·FEMS Immunology and Medical Microbiology·S FournierF Derouin
Nov 29, 2001·Journal of Clinical Microbiology·B DengjelH Rinder
Mar 7, 2002·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·Beatriz LoresCarmen del Aguila
Aug 22, 2002·The Journal of Eukaryotic Microbiology·William V EversonH D Alan Lindquist

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Citations

Feb 2, 2010·Emerging Infectious Diseases·Bohumil SakEvan W Secor
Feb 21, 2014·Parasitology Research·J G Sánchez ColladoR Martín-Hernández

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