Luminol reagent and acridine ester reagent are commonly used in chemiluminescence immunoassay. Chemiluminescence immunoassay has replaced enzyme-linked immunosorbent (ELISA) as a more mainstream in vitro diagnostic method, accounting for more than 50% of biochemical detection methods.
The attraction of chemiluminescence immunoassay as an in vitro diagnostic technique lies in its simplicity. The essence of chemiluminescence is self luminescence, which means that the analytical instrument of chemiluminescence only needs to provide a method to detect the light signal and record the results. The autoluminescence detector needs a closed light sample chamber and a photo detector. The simplest is photo paper or X-ray, or even a visual detector.
The convenience of chemiluminescence detection method makes its application simple and completely automatic, but what is its sensitivity? Chemiluminescence has two inherent advantages.
1. Most of the samples have no background signal, such as they do not emit light.
2. Chemiluminescence detection is not a proportional test, which is different from fluorescence and absorption or colorimetric test. In the fluorescence test, it is very difficult to detect the fluorescent group with small Stokes shift, and the fluorescence is difficult to distinguish from the excitation wavelength.
Another problem is that some stray light will enter the detector, especially when the sample is turbid. In the absorbance test, the fundamental factor limiting the absorbance is to distinguish a small difference between two relatively strong signals.
It should be noted that the sensitivity of the detector to the spectrum of chemiluminescence is as close as possible, and the maximum sensitivity has been obtained. In general, the photomultiplier tube in autoluminescence instrument has the best response to blue light, and is not sensitive to the end spectrum of red light. The solid state detector has a good response to red light.
X-ray films are widely used for chemiluminescence imprinting analysis on nylon, cellulose or PVDF membranes. But we need to keep in mind that X-ray films are only used to detect light signals in the spectrum from ultraviolet to blue, although there are some special films that are sensitive to enhanced green light.
Chemiluminescence detection includes liquid samples and solid samples. The indicators of chemiluminescence detection are usually sensitivity, linearity and dynamic range. Desheng's chemiluminescence reagents include luminol, isoluminol and six acridine ester substrates.
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