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What are the classifications of chemiluminescent reagents
Chemiluminescence immunoassay (CLIA) was born in 1977 and is more advanced than many traditional detection methods. This technology has high sensitivity, wide linear range and wide application range. Moreover, compared with radioimmunoassay (RIA), fluorescence immunoassay (IFA) and enzyme-linked immunoassay (EIA), it is easy to operate, fast in response, no radioactive pollution, simple and stable instrument and can be automated.
Chemiluminescence is the light radiation produced when the reactant or product of a chemical reaction absorbs the chemical energy released by the reaction, and the electron transitions from the ground state to the excited state, and then returns from the excited state to the ground state. Chemiluminescence behavior is often determined by the chemical properties of the oxidant And spectral properties.
According to the luminous time, it can be divided into two types: Flash and Glow:
1. The luminescence time is within a few seconds. For example, the luminescence time of acridinium ester is usually only about 2 seconds, which is measured by in-situ injection (In Situ Injector) and time integration method. The chemiluminescence of acridinium ester belongs to the flash type.
2. The glow time is more than tens of minutes, such as: horseradish peroxidase plus luminol system, alkaline phosphatase plus AMPPD system, xanthine oxidase system, no need for in-situ sample injection, with speed Method measurement.
The solid phase carrier of the chemiluminescent immune system is divided into two types: plate type and tube type:
Luminol, a chemiluminescent reagent, emits a blue glow when oxidized
1. Plate chemistry is the immune system in which the solid phase carrier is the microplate, which requires a long reaction time.
2. Tubular chemiluminescence refers to the immune system in which the solid-phase carrier is microparticles, which is generally referred to as magnetic beads or microspheres. Most of the tube chemiluminescence is a large-scale immunoassay system, and this advanced tube chemiluminescence method has been used in POCT in China.
Different chemiluminescence reagents use different chemiluminescence systems:
1. Direct chemiluminescence: Acridinium ester is oxidized by hydrogen peroxide under alkaline conditions and can flash quickly. Acridinium ester derivatives have high labeling efficiency and good stability. Therefore, acridinium ester is used as a marker for immunoassay and luminescence The system is simple, low-cost, has a low background and can be quickly detected.
2. Enzymatic chemiluminescence: Enzyme-labeled biologically active substances are used for immunoreaction. The enzymes on the immunoreaction complex then act on the luminescent substrate to emit light under the action of the signal reagent, and perform luminescence measurement with a luminescence signal analyzer. Commonly used are HRP-H2O2-LUMINOL (luminol) system and ALP-AMPPD luminescence system. Luminol is oxidized by strong oxidants to emit light without enzyme catalysis.
Electrochemiluminescence immunoassay and photo-induced chemiluminescence (LiCA) are more advanced methods, and they are currently not used in China. Therefore, the chemiluminescence reagents currently produced by Desheng are based on luminol, isoluminol, acridine esters and their Mainly derivatives, product performance can replace imported products.