Description
The Eagle Biosciences specimens are pipetted into wells of a microtiter plate previously coated with antibodies directed against HAV. The HAV Antigens binds to the fixed antibody and after the incubation period of two hours at 37°C the plate is washed thoroughly. Bound HAV Antigens is identified by conjugate addition (monoclonal anti-HAV, peroxidase conjugated) incubated for another two hours at 37°C. Excess conjugate is removed by washing and the substrate is added. After 30 minutes incubation at room temperature the reaction is terminated by adding stop solution. The blue colour of a positive reaction turns to yellow and is measured in a microplate reader at 450 nm. The intensity of the colour indicates the concentration of bound HAV Antigens. A positive reaction must be confirmed by neutralising with anti-HAV serum to discriminate false positive reactions which sometimes occurs in stool