The principle of the following enzyme immunoassay test follows a typical two-step capture or ‘sandwich’ type assay.
The assay makes use of two highly specific monoclonal antibodies: A monoclonal antibody specific for CRP is immobilized onto the microwell plate and another monoclonal antibody specific for a different region of CRP is conjugated to horse radish peroxidase (HRP).
CRP from the sample and calibrators are allowed to bind to the plate, washed, and subsequently incubated with the HRP conjugate.
After a second washing step, the enzyme substrate is added.
The enzymatic reaction is terminated by addition of the stopping solution.
The absorbance is measured on a microtiter plate reader.
The intensity of the colour formed by the enzymatic reaction is directly proportional to the concentration of CRP in the sample.
A set of calibrators is used to plot a calibration curve from which the amount of