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Diluted patient serum (serum diluent contains sorbent to remove rheumatoid factor and humanIgG interference) is added to wells coated with purified antigen. IgM specific antibody, if present,binds to the antigen. All unbound materials are washed away and the enzyme conjugate isadded to bind to the antibody-antigen complex, if present. Excess enzyme conjugate is washedoff and substrate is added. The plate is incubated to allow the oxidation of the substrate by theenzyme. The intensity of the color generated is proportional to the amount of IgM specificantibody in the sample.