Immunomagnetic separation of Toxoplasma gondii oocysts using a monoclonal antibody directed against the oocyst wall.
Recent outbreaks of waterborne toxoplasmosis have stimulated the development of sensitive methods to detect Toxoplasma gondii oocysts in samples suspected to be contaminated. The immunomagnetic separation (IMS) have been standardised to detect waterborne protozoa, but it did not exist for Toxoplasma oocysts. In this study, we describe two monoclonal antibodies (mAbs 3G4 and 4B6) produced against the oocyst wall, and the incorporation of mAb 3G4 in an IMS procedure. We found that an indirect IMS method gave better mean recoveries than a direct one (69.4% and 25.2%, respectively). Dissociation of oocyst_magnetic bead complexes was greatly improved by using a 2% aqueous H2SO4 solution instead of a 0.1 N HCl solution (82.8% and 17.4%, respectively). With these parameters, mean recoveries of less than 1000 oocysts ranged from 44.6% to 82.9%, depending on incubating temperature and buffer. Age of oocysts (1 or 12 months old) does not influence IMS performances. Results of this study indicate that the described IMS is an efficient technique to recover Toxoplasma oocysts.
Dumètre
Aurélien
A
UPRES EA3174, Neuroparasitologie et Neuroépidémiologie Tropicale, Faculté de Médecine, 2 Rue du Dr Marcland, F-87025 Limoges, France.
Dardé
Marie-Laure
ML
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000737:chemistry* / Q000276:immunology; D002473:Cell Wall / Q000276:immunology; D005260:Female; D005455:Fluorescent Antibody Technique; D006801:Humans; D018189:Immunomagnetic Separation / Q000379:methods*; D051379:Mice; D008863:Microspheres; D033661:Oocysts / Q000302:isolation & purification; D014122:Toxoplasma / Q000166:cytology / Q000302:isolation & purification*; D014123:Toxoplasmosis / Q000469:parasitology* / Q000517:prevention & control; D014881:Water Supply
D000911:Antibodies, Monoclonal