Capture method. Total duration of assay: 70 minutes
This kit uses capture ELISA principle to detect CMV IgM. Polystyrene microwell
strips are pre-coated with mouse anti-human IgM (anti-µ chain specific). During
the first incubation step, all IgM antibodies binds to mouse anti-human IgM
monoclonal antibody coated in microplate including the Anti-CMV IgM, the wells
are washed to remove unbound serum proteins, and CMV antigens conjugated
to the enzyme horseradish peroxidase (HRP-Conjugate) are added. During the
second incubation step, if there is CMV IgM antibody in sample, HRP
conjugated antigens will be bound to the anti-human CMV IgM complexes
previously formed and the unbound HRP-conjugate is then removed by washing.
Chromogen solutions containing Tetramethylbenzidine (TMB) and urea peroxide
are added to the wells and in presence of the anti-human IgM -antibody(IgM
antigen (HRP) immunocomplex, the colorless Chromogens are hydrolyzed by
the bound HRP conjugate to a blue-colored product. The blue color turns yellow
after stopping the reaction with sulfuric acid.
The amount of color intensity can be measured and it is proportional to the
amount of antibody captured in the wells, and to the amount of antibody in the
sample respectively