The Leptospira IgM-ELISA is intended for the qualitative determination of IgM class antibodies against Leptospiraspp. in human serum or plasma (heparin).
The qualitative immunoenzymatic determination of IgM-class antibodies against Leptospira spp. is based on the ELISA (Enzyme-linked Immunosorbent Assay) technique. Microtiter strip wells are coated with Leptospira antigens to bind corresponding antibodies of the specimen. After washing the wells to remove all unbound sample material horseradish peroxidase (HRP) labelled anti-human IgM conjugate is added. This conjugate binds to the captured Leptospira-specific antibodies. The immune complex formed by the bound conjugate is visualized by adding tetramethylbenzidine (TMB) substrate which gives a blue reaction product. The intensity of this product is proportional to the amount of Leptospira-specific IgM antibodies in the specimen. Sulfuric acid is added to stop the reaction. This produces a yellow endpoint color. Absorbance at 450nm is read using an ELISA microwell plate reader.