<P> Antigens from the sample are attached to a surface . Then, a further specific antibody is applied over the surface so it can bind to the antigen . This antibody is linked to an enzyme, and, in the final step, a substance containing the enzyme's substrate is added . The subsequent reaction produces a detectable signal, most commonly a color change in the substrate . </P> <P> Performing an ELISA involves at least one antibody with specificity for a particular antigen . The sample with an unknown amount of antigen is immobilized on a solid support (usually a polystyrene microtiter plate) either non-specifically (via adsorption to the surface) or specifically (via capture by another antibody specific to the same antigen, in a "sandwich" ELISA). After the antigen is immobilized, the detection antibody is added, forming a complex with the antigen . The detection antibody can be covalently linked to an enzyme, or can itself be detected by a secondary antibody that is linked to an enzyme through bioconjugation . Between each step, the plate is typically washed with a mild detergent solution to remove any proteins or antibodies that are non-specifically bound . After the final wash step, the plate is developed by adding an enzymatic substrate to produce a visible signal, which indicates the quantity of antigen in the sample . </P> <P> Of note, ELISA can perform other forms of ligand binding assays instead of strictly "immuno" assays, though the name carried the original "immuno" because of the common use and history of development of this method . The technique essentially requires any ligating reagent that can be immobilized on the solid phase along with a detection reagent that will bind specifically and use an enzyme to generate a signal that can be properly quantified . In between the washes, only the ligand and its specific binding counterparts remain specifically bound or "immunosorbed" by antigen - antibody interactions to the solid phase, while the nonspecific or unbound components are washed away . Unlike other spectrophotometric wet lab assay formats where the same reaction well (e.g. a cuvette) can be reused after washing, the ELISA plates have the reaction products immunosorbed on the solid phase which is part of the plate, and so are not easily reusable . </P> <P> As an analytic biochemistry assay, ELISA involves detection of an "analyte" (i.e. the specific substance whose presence is being quantitatively or qualitatively analyzed) in a liquid sample by a method that continues to use liquid reagents during the "analysis" (i.e. controlled sequence of biochemical reactions that will generate a signal which can be easily quantified and interpreted as a measure of the amount of analyte in the sample) that stays liquid and remains inside a reaction chamber or well needed to keep the reactants contained; It is opposed to "dry lab" that can use dry strips--and even if the sample is liquid (e.g. a measured small drop), the final detection step in "dry" analysis involves reading of a dried strip by methods such as reflectometry and does not need a reaction containment chamber to prevent spillover or mixing between samples . </P>

Elisa test is used to detect which of the following disease