Anti-HAV ELISA

The Anti-HAV ELISA is a pseudo-competitive enzyme immunoassay. Serum or plasma samples are added to the wells of a microplate, which has been previously coated with inactivated HAV antigen, and incubated for 2 hours at 37°C. Anti-HAV antibodies bind to the antigen. The conjugate (peroxidase labeled anti-HAV) is added and incubated again for 1 h at 37°C. Free binding sites of the antigen are bound with conjugate. Excess conjugate is washed off the plate and the substrate is added and incubated for 30 min at room temperature. The bound conjugate changes the color of the subtrate to blue. The reaction is terminated by adding the stop solution. The color turns yellow. The absorbance of the colored reaction product is measured on a microtiter plate reader. The absorbance is reciprocal to the anti-HAV titer. For quantitative determination use the included serum standard for preparation of a titration curve.