Estandarización del inmunoensayo ELISA para la detección de IgG anti-Toxoplasma gondii en ratón

Abstract

We describe a standardization of an enzyme-linked immunosorbent assay (ELISA) for detection of specific anti-Toxoplasma gondii IgG in mice. This assay is important as a tool for strains characterization in T. gondii and for diagnosis of human toxoplasmosis. The technique was standardized by testing different conditions (different coating reagents and different amounts of antigen). No significant difference was observed between coating reagents; therefore buffer coating with bovine albumin 1% was chosen. The best amount of antigen was 20 μg/ml at a 1:1000 dilution of the secondary antibody. The cut-off was the average value of all negative serum + 2 deviation standard (0,135). Negative serum values were below of cut-off at 410 nm. We do not found differences in detection among strains of T. gondii. These results show that ELISA is highly sensitive and specific and is useful to improve efficiency in procedures that involve animals in the laboratory.