Enzyme ImmunoAssay (ELISA) for the qualitative determination of IgM antibodies to Dengue Virus in human plasma and sera.

Micro-plates are coated with higly purified Dengue Virus antigen. In the 1st incubation, the solid phase is treated with diluted samples and anti Dengue Virus IgM are captured, if present, by the antigens. After washing out all the other components of the sample, in the 2nd incubation bound anti Dengue Virus IgM are detected by the addition of anti hIgM antibody, labeled with peroxidase (HRP).The enzyme captured on the solid phase, acting on the substrate/chromogen mixture, generates an optical signal that is proportional to the amount of anti Dengue Virus IgM antibodies present in the sample. Neutralization of IgG anti-DV, carried out directly in the well in the 1st incubation, is performed in the assay in order to block interferences due to this class of antibodies in the determination of IgM.