How does PCR-Elisa work?

The process begins when the DNA sequence of interest is amplified using PCR. During this process, digoxigenin-11-dUTP (DIG-dUTP) is incorporated into the DNA, labeling the PCR products. The amplified DNA is tagged with biotin and the biotinylated DNA is then immobilized onto a microplate (often a 96-well plate) that has been coated with streptavidin. After immobilization, the presence of the target DNA is detected using specific antibodies. For example, an anti-DIG antibody linked to the enzyme peroxidase is added. These antibodies are typically linked to an enzyme that produces a color change or a fluorescent signal when a substrate (e.g. ABTS) is added. The amount of color or fluorescence is then measured using a spectrophotometer or a plate reader.