This method is particularly useful when it's difficult to remove interfering substances from the antigen or when obtaining a sufficient amount of purified antigen is challenging. The principle behind this technique involves competition between the antibodies in the sample and a known amount of enzyme-labeled antibody for binding to the solid-phase antigen. The higher the concentration of antibodies in the specimen, the fewer enzyme-labeled antibodies will bind to the solid phase, resulting in a weaker positive reaction compared to the negative control. If the antigen is highly pure, it can be directly coated onto the solid phase. However, if there are impurities present, direct coating may not be effective. In such cases, a capture coating method is often used. This involves first immobilizing an antibody that specifically recognizes the antigen onto the solid phase, followed by adding the antigen to form a complex. Any impurities in the antigen are then removed through washing before the sample and enzyme-labeled antibody are added for the competitive binding reaction. There are several variations of the competitive assay. One common approach is to allow the sample and the enzyme-labeled antibody to compete for binding to the solid-phase antigen. This method is widely used in anti-HBc ELISA. Another variation involves mixing the sample with the antigen and adding them to the solid-phase antibody for initial binding. After washing, the enzyme-labeled antibody is introduced to react with the antigen that has been captured on the solid phase. This method is typically employed for detecting anti-HBe. By using these different competitive formats, laboratories can effectively detect various types of antibodies, even in complex or impure samples, ensuring accurate and reliable results in diagnostic testing.
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