Development of a Rapid Solid Phase Immunoassay for the Detection of Flavivirus Antigens in Human Blood and Mosquito Vectors

摘要

Antigen-capture solid phase immunoassays employing flavivirus monoclonal antibodies (MAB) for the detection of yellow fever (YF) or dengue (DEN) viruses in cell cultures and serum have recently been reported. The assays, which employs virus type-specific, complex-reactive or group-reactive MAB in the capture phase, are reasonably sensitive and specific. The goal of our work has been the development of a generic flavivirus detection system and emphasis has been placed on an enzyme-linked immunoassay (ELISA) for the detection of DEN-infected mosquitoes. Attention has been directed at the identification and characterization of MAB with broad flavivirus group-reactive domain (as determined by competitive binding assay) with the expectation that such antibody combination will provide the best opportunity for antigen binding and therefore maximum assay sensitivity. However, since the available broadly group-reactive MAB has been found to exhibit at least some degree of cross-competition, flavivirus type-specific detector MAB directed at sites distant from the group-reactive domain have been extensively used to evaluate the properties of potential flavivirus group-reactive capture antibodies. Initial assays demonstrated the capacity of the YF-derived antibodies to capture prototype flaviviruses.