The Leptospira IgM ELISA test is a qualitative enzyme immunoassay for the detection of antibodies to Leptospira biflexa in samples of human serum or plasma. The microwells are coated with purified Leptospira biflexa antigen. During the first incubation with the diluted patients' sera, any antibodies which are reactive with the antigen will bind to the coated wells. After washing to remove the rest of the sample, the Enzyme Conjugate is added. If antibodies have been bound to the wells, the Enzyme Conjugate will then bind to these antibodies. After another series of washes, a chromogen (tetramethylbenzidine, or TMB) is added. If the Enzyme Conjugate is present, the peroxidase will catalyze a reaction that consumes the peroxide and turns the chromogen from clear to blue. Addition of the Stop Solution ends the reaction and turns the blue color to a bright yellow color. The reaction may then be read visually or with an ELISA reader.