Efficient duplex solid-phase fluorescent assay (dFISA) for the simultaneous detection of specific anti-T. gondii IgG and IgM due to refined conjugates

Toxoplasma gondii infections are very common, causing occasional central nervous system and eye diseases, and must be screened in prenatal care for efficient therapy. Here, we developed a duplex solid-phase fluorescent assay (dFISA) for the simultaneous detection of anti-T. gondii IgG and IgM antibodies in prenatal care screening for toxoplasmosis. Assays using commercially available ion-exchange purified conjugates yielded poor results and high background fluorescence. Same-well IgG/IgM dFISA with refined conjugates was used to test 140 samples from university students, 120 samples from pregnant women and 24 samples from adult volunteers at a large public hospital. We found that dFISA offers high concordance, specificity and reproducibility for IgG (Kappa = 0.883) and IgM (Kappa = 0.918), which is useful in high-throughput applications for antenatal care.