Method Description

Polystyrene microwells are coated with the antihuman antibody specific for IgM (mu-chain). Diluted serum specimens and controls are incubated in the wells. The IgM present in the specimen binds to the antihuman antibody (IgM specific) in the wells. Nonspecific reactants are removed by washing. WNV antigen is then added to the wells and incubated. If anti-WNV IgM is present in the specimen, the WNV antigen binds to the anti-WNV in the well. Unbound WNV antigen is then removed by washing the well. Mouse anti-flavivirus conjugated with horseradish peroxidase (HRPO) is then added to the wells and incubated. If WNV antigen has been retained in the well by the anti-flavivirus in the specimen, the mouse anti-flavivirus:HRPO binds to WNV antigen in the wells. Excess conjugate is removed by washing. Enzyme substrate and chromogen are added, and the color is allowed to develop. After adding the Stop reagent, the resultant color change is quantified by a spectrophotometric reading of optical density (OD) that is directly proportional to the amount of antigen-specific IgM present in the specimen. Specimen OD readings are compared with reference cutoff OD readings to determine results.(Package insert: West Nile Virus IgM Capture DxSelect. Focus Diagnostics; 05/08/2018)