Development of an Assay for detection of IgM to Hantaan Virus Using Serum Bound to Porous Membranes to Entrap Antigen Coated Latex Beads

Abstract

A novel immunoassay to detect IgM antibodies to Hantaan virus (family Bunyaviridae) in human sera was developed. The most unique feature of this rapid diagnostic immunoassay was the substitution of colored latex beads in place of enzymesubstrate reactions. Viral antigen-coated latex beads were entrapped in porous membranes (as they were pulled through under vacuum) coated with anti-human IgM serum which specifically captured IgM type antibodies. Development of a model system using mouse sera and antiserum to mouse IgG preceded development of the Hantaan immunoassay. The model system exhibited distinctive and reproducible specific entrapment of antigen coated beads within the membrane matrix. Although the immunological and experimental variables within this immunoassay were carefully controlled and analyzed, the assay for Hantaan IgM antibody was not diagnostically reliable due to the fact that their was not enough resolution between positive results and negative controls to be visually discernible.