Method Description

This enzyme-linked immunosorbent assay is designed to detect IgM and IgG class antibodies to Borrelia burgdorferi in human sera. The sensitized wells of plastic microwell strips are prepared by passive adsorption with B burgdorferi whole cell antigen. The test procedure involves 3 incubations steps. First, test sera (properly diluted) are incubated in antigen coated microwells. Any antigen-specific antibody in the sample will bind to the immobilized antigen. The plate is washed to remove unbound antibody and other serum components. Second, peroxidase conjugated goat anti-human IgM (mu chain specific) and IgG (Fc chain specific) is added to the wells, and the plate is incubated. The conjugate will react with IgM and IgG antibody immobilized on the solid phase in the first step. The wells are washed to remove unreacted conjugate. Third, the microwells containing immobilized peroxidase conjugate are incubated with peroxidase substrate solution. Hydrolysis of the substrate by peroxidase produces a color change. After a period of time, the reaction is stopped, and the color intensity of the solution is measured photometrically. The color intensity of the solution depends upon the antibody concentration in the original sample.(Package insert: B burgdorferi IgM or IgG Test System. ZEUS Scientific, Inc; Rev Date 01/27/2020)