Toxoplasma gondii: purification of zoites from peritoneal exudates by eight methods.
Many methods have been proposed for removing contaminating host cells from mouse peritoneal exudates infected with Toxoplasma gondii tachyzoites. Of these, eight established methods were compared. They were density gradients, sonication and trypsin digestion, differential centrifugation, haemolysin digestion, filtration through glass wool and cellulose columns, and sintered glass and polycarbonate filtration. The methods were assessed for zoite recovery, host cell removal, effect on zoite viability and antigenic integrity, time, cost, and ease. They were almost all capable of removing greater than 90% of the mouse leucocytes, but in some cases this resulted in low zoite recoveries. The sonication and trypsin method produced the best zoite recovery and highest purity, but appeared to affect zoite viability and antigenic integrity. The haemolysin digestion procedure has been adopted by our laboratory because of its high recovery of zoites, and it is inexpensive, quick, and easy to perform.
D000818:Animals; D000941:Antigens / Q000276:immunology; D001202:Ascitic Fluid / Q000469:parasitology*; D002498:Centrifugation; D002499:Centrifugation, Density Gradient; D003365:Costs and Cost Analysis; D005260:Female; D005374:Filtration; D006460:Hemolysin Proteins; D007962:Leukocytes; D051379:Mice; D009504:Neutrophils; D013010:Sonication; D014122:Toxoplasma / Q000276:immunology / Q000302:isolation & purification* / Q000502:physiology; D014124:Toxoplasmosis, Animal / Q000469:parasitology*; D014357:Trypsin
D000941:Antigens; D006460:Hemolysin Proteins; D014357:Trypsin