Description
The principle of the following enzyme immunoassay test follows a two-step competitive binding scenario. Competition occurs between an unlabelled antigen (present in standards, controls and patient samples) and a biotin-labelled antigen (conjugate) for a limited number of antibody binding sites on the microplate. In the second step, the streptavidin-horseradish peroxidase conjugate binds to any bound biotinylated testosterone. The washing and decanting procedures remove unbound materials. After the last 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 is inversely proportional to the concentration of testosterone in the sample. A set of standards is used to plot a standard curve from which the amount of testosterone in patient samples and controls can be directly read