Intended use of the test kit
The principle of the test is based on the reaction of 2 monoclonal antibodies with 2 different antigenic determinants on the gE glycoprotein of Aujeszky’s disease virus (ADV). Where a negative sample does not block the reaction, a sample containing antibodies to gE does block the reaction. However, pigs immunized with vaccines lacking gE expression do not block the reaction and thus are scored negative. In this way naturally, infected pigs can be discriminated from vaccinated pigs.
Principle of the test kit
A first monoclonal anti-gE antibody (the catching antibody) is used for coating the wells of a 96-well microtiter plate. Test sample and antigen, consisting of ADV infected cell cultures, are incubated in the microtiter test plate after 2 hours. A second monoclonal anti gE antibody, conjugated with horseradish peroxidase (HRPO) is added to the wells. This monoclonal antibody recognizes a different antigenic determinant on gE compared to the catching antibody.
The sample is added (diluted 1:1) to the wells of the coated plate.
The sample also can be titrated using a 3-step dilution, starting with a dilution 1:1(>1:3 >1:9 >1:27).
After incubation and washing, the substrate is added. If the test sample is negative, i.e. does not contain antibody to gE, HRPO and substrate will produce a colour reaction. If the test sample is positive, the binding of the antigen to one or both monoclonal antibodies will be blocked and the colour reaction fails to appear. A test sample is defined positive if the extinction is below 68% of the mean of that of the negative control which is included in this test kit.