Development of an Experimental ELISA for the Detection of Antigens and Antibodies to the Virus of Pacheco Disease using Monoclonal Antibodies

Abstract

Monoclonal antibodies (Mabs) against one of the strains of Pacheco Parrot's Disease (PPD) were produced and characterized. An enzyme-linked immunosorbent assay (ELISA) for the rapid detection of antigen and antibody to PPD was developed. The antigen ELISA could detect up to 11 7 ng/ml of purified virus material. To overcome the requirement for large amounts of purified viral antigen, the microtitre trays were initially coated with Mabs prepared against purified PPD virus (PPDV). A relatively crude viral preparation could then be used to coat the trays. Using one more step of incubation between the viral preparation and a test sera, the same ELISA protocol could be used for screening antibody against PPDV. The blocking ELISA with a detection range of 625ng