Luminol is a kind of chemical fluorescence molecule. In the presence of hydrogen peroxide molecule, luminol can be converted into excited amino-o-phthalic acid, which emits strong fluorescence. Hydrogen peroxide is the product of many biological oxidation reactions, so it is easy to associate these biological oxidation reactions with light detection by introducing luminol. For example, the glucose oxidase / catalase probe can detect the concentration of hydrogen peroxide or glucose in the sample, and the response time is only 0.5s (dynamic method). Chemiluminescent immunoassay (CLIA) is a technique that combines chemiluminescent substances with immunological reaction and expresses the concentration of the tested immune components by light reaction. In 1976, shmeder put forward chemiluminescent immunoassay, using luminol-h202 and its derivative Abei as the detection reaction display system.

At present, luminol and its derivatives have been the most commonly used markers in the application of chemiluminescent immunoassay. They can release a large number of photons under the alkaline condition through the catalysis of microperoxidase Nordic acid can be used for hybridization detection of HRP labeled antibody with Western blot and HRP labeled probe, and also for detection of modern criminal blood trace. The compounds that can be used as chemiluminescent markers must have the following conditions: high quantum yield of luminescence; its physical and chemical properties match the studied system; its luminescence reaction is the result of the oxidation reaction of luminescent substances; it is not toxic to organisms within the concentration range used. Some commonly used chemiluminescent reagents are acridiniumester, which is a kind of widely used chemiluminescent tracer. It is a tricyclic organic compound, easy to oxidize, and no catalyst is needed for the oxidation reaction, and photons are released at 430nm.

Luminol and isoluminol and their derivatives are the most mature chemiluminescent agents, which were studied by white as early as 1964 It is reported that luminol, isoluminol and their derivatives have been used in chemiluminescent immunoassay. Luminescence detection: the optimal fluorescence wavelength is 400nm (detect the chemiluminescence in 60mmk2s2o8100mmk2co3, ph11.5 solution)