Flow cytometric application of the Sabin and Feldman dye test in the diagnosis of toxoplasmosis.
Although time-consuming and requiring live parasites, the Sabin and Feldman dye test (DT) is still considered the 'gold standard' among the serological tests for toxoplasmosis diagnosis. The present study was initiated to compare detection of dead parasites using optical microscopy with flow cytometry and a fluorescent nonvital dye, propidium iodide. After incubation with sera (N = 150) and a complement source, tachyzoites were washed, then stained using a fluorescein-conjugated Toxoplasma-specific antiserum. Dead tachyzoites were detected by flow cytometry after addition of propidium iodide. Intra- and inter-assay reproducibilities of percentages of dead parasites varied between 7 and 14%, and 8 and 21%, respectively. When comparing flow cytometry with the classical DT, no discrepancy was noted for positive (N = 118) and negative sera (N = 32). Correlation was good (r = 0.85) for positive sera. In conclusion, when easily available, flow cytometry is a very sensitive, specific and time-sparing method to detect specific antibodies to Toxoplasma gondii.
Cozon
G J
GJ
Unité d'Immunologie, Hôpital de la Croix-Rousse, Lyon, France. gcozon@rockefeller.univ-lyon1.fr
Ferrandiz
J
J
Thulliez
P
P
Peyron
F
F
D000818:Animals; D005260:Female; D005434:Flow Cytometry / Q000379:methods; D005456:Fluorescent Dyes; D006801:Humans; D011247:Pregnancy; D011419:Propidium; D015203:Reproducibility of Results; D014123:Toxoplasmosis / Q000175:diagnosis*
D005456:Fluorescent Dyes; D011419:Propidium