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Isolation of Purified Oocyst Walls and Sporocysts from Toxoplasma Gondii

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Abstract:Toxaplasma gondii is a parasitic protozoan that infects a wide range of vertebrates, including humans. This report describes methods that have been developed for separation of oocyst components starting with the mechanical fragmentation of oocysts. Use of iodixoanol gradients and centrifugation gave the highest yield of relatively pure fractions enriched in oocyst walls and intact sporocysts. Percoll gradient centrifugation also resulted in fractions highly enriched in intact sporocysts. Flow Cytometry with fluorescence activated cell sorting (FACS) was useful for analytical purposes. FACS yielded analytical quantities of oocyst walls and intact sporocysts. Flow cytometry with FACS also proved useful for quantitation of purity obtained following iodixanol gradient fractionation. These novel methods for isolation of oocyst and sporocysts provide materials that have utility in many applications. Oocyst walls may be used as an enriched starting material for development of methods to detect the organism in environmental samples, sporocysts as a starting material for analysis of excystation, and use of either fraction for proteomic analysis of components unique to this life cycle stage.
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Citation:Everson, W. V., M. W. Ware, and H. A. Lindquist. Isolation of Purified Oocyst Walls and Sporocysts from Toxoplasma Gondii. JOURNAL OF EUKARYOTIC MICROBIOLOGY 49(4):344-349, (2002).
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Contact: Mary P. O'Bryant - (919)-541-4871 or obriant.mary@epa.gov
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Division: Microbiological & Chemical Exposure Assessment Division
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Branch: Biohazard Assessment Research Branch
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Product Type: Journal
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Published: 07/01/2002
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Related Entries:
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Bullet Item Detecting Ccl-Related, Emerging and Regulated Waterborne Human Protozoa for Exposure Assessment
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Last Updated on Monday, October 22, 2007
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