Description
The anti-HAV IgM EIA 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.
