Chlamydia Trachomatis IgM ELISA

The Chlamydia trachomatis IgM ELISA Kit is a solid-phase enzyme-linked immunosorbentassay (ELISA). Samples are diluted with Sample Diluent and additionally incubated with IgG-RFSorbent, containing hyper-immune anti-human IgG-class antibody to eliminate competitiveinhibition from specific IgG. This pretreatment avoids false negative results. Microtiter wells as asolid phase are coated with inactivated Elementary Bodies Chlamydia trachomatis antigen.Diluted samples and ready-to-use controls are pipetted into these wells. During incubationChlamydia trachomatis-specific antibodies of positive samples and controls are bound to theimmobilized antigens. After a washing step to remove unbound sample and control materialhorseradish peroxidase-conjugated anti-human IgM antibodies are dispensed into the wells.During a second incubation, this anti-IgM conjugate binds specifically to IgM antibodies resultingin the formation of enzyme-linked immune complexes. After a second washing step to removeunbound conjugate, the immune complexes formed (in case of positive results) are detected byincubation with TMB substrate and development of a blue color. Addition of sulfuric acid stops theenzymatic reaction and turns the solution from blue to yellow. The intensity of this color is directlyproportional to the amount of Chlamydia trachomatis-specific IgM antibody in the sample.Absorbance at 450 nm is read using an ELISA microtiter plate reader.