Purified HSV-1 antigen is coated on the surface of microwells. Diluted patient serum is added to the wells, and the HSV-1 IgM- specific antibody, if present, binds to the antigen. All unbound materials are washed away. HRP-conjugate is added, which binds to the antibody-antigen complex. Excess HRP-conjugate is washed off and a solution of TMB Reagent is added. The enzyme conjugate catalytic reaction is stopped at a specific time. The intensity of the color generated is proportional to the amount of HSV-1 IgM-specific antibody in the sample. The results are read by a microwell reader compared in a parallel manner with calibrator and controls