AN IMPROVED ASSAY FOR THE DETECTION OF TOXOPLASMA-GONDII ANTIBODIES IN HUMAN SERUM BY FLOW-CYTOMETRY

The diagnosis of toxoplasmosis relies primarily on the demonstration o f specific antibodies to Toxoplasma gondii. We describe a flow cytomet ry method for the determination of antibodies to whole fixed tachyzoit es by indirect immunofluorescence. Fixed tachyzoites in suspension hav e characteristic light scattering properties. The amount of IgM, IgG, and IgA antibodies from patients' sera bound to the tachyzoites can be estimated from the mean fluorescence intensities observed using class -specific conjugates for different Ig heavy chains. Appropriate serum dilutions for the estimation of specific IgG titres and for the discri mination between sera positive and negative for IgA and IgM antibodies were established using 40 random sera from pregnant women. The method proved to be quantitative and highly sensitive as compared with curre ntly used assays. Coefficients of variation between series ranged from 6.4% to 12.2% and could be controlled by the inclusion of positive an d negative standard sera in the test series. The method might represen t a major improvement in cost effectiveness as compared with routine i mmunofluorescence assay by fluorescent microscopy when the volume of t esting is sufficient. It can be fully automated, and its large scale a pplication is currently under investigation.