The Eagle Biosciences Anti-HAV IgM ELISA Assay Kit is a class-capture enzyme immuoassay. Serum or plasma samples are diluted 1:2000 and added to the wells of a microtiter plate, which have been previously coated with an antibody directed against human IgM antibodies (μ-chain specific). Specific antibodies in the sample bind to the antibodies present during an incubation step of 1h at 37°C. After washing, the Hepatitis A-virus is added. During an incubation step of 1h at 37°C the HAV binds to the HAV specific IgM antibodies. The conjugate (monoclonal anti-HAV antibody, peroxidase-labelled) is added and incubated again (for 1 h at 37°C). After a further washing step substrate is added and further incubated for 30 min. at room temperature. The reaction is terminated on addition of stop solution. The absorbance of the coloured reaction product is measured on a microtiter plate reader. The colour intensity of the reaction corresponds to the concentration of antibodies in the sample. Sample Types: serum, plasma. Time: 3.5 hours ? Positive detection of antibodies directed against the Hepatitis A virus (anti-HAV) is evidence of immunity to Hepatitis-A virus (1). After natural infection with the Hepatitis A-Virus, neutralising antibodies appear at the same time of Anti-HAV of IgG-Class formation (1).