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The DRG Toxoplasma gondii IgM Enzyme Immunoassay Kit is for determination of IgM-class antibodies to Toxoplasma gondii in serum.
The DRG Toxoplasma gondii IgM ELISA Kit is a solid phase enzyme-linked immunosorbent assay (ELISA). Samples are diluted with Sample Diluent and additionally incubated with IgG-RF-Sorbent, containing hyper-immune antihuman IgG-class antibody to eliminate competitive inhibition from specific IgG and to remove rheumatoid factors. This pretreatment avoids false negative or false positive results.
Microtiter wells as a solid phase are coated with detergent treated Toxoplasma gondii antigen (Deelen strain). Pretreated specimens, ready-for-use standards and ready-for-use control are pipetted into these wells. During incubation Toxoplasma gondii-specific antibodies of positive specimens, and standards are bound to the immobilized antigens.
After a washing step to remove unbound sample, standard and control material horseradish peroxidase conjugated antihuman IgM antibodies are dispensed into the wells. During a second incubation this anti-IgM conjugate binds specifically to IgM antibodies resulting in the formation of enzyme-linked immune complexes.
After a second washing step to remove unbound conjugate the immune complexes formed (in case of positive results) are detected by incubation with TMB substrate and development of a blue color. The blue color turns into yellow by stopping the enzymatic indicator reaction with sulfuric acid.
The intensity of this color is directly proportional to the amount of Toxoplasma gondii-specific IgM antibody in the specimen. Absorbance at 450 nm is read using an ELISA microtiter plate reader.