Chemiluminescent kit for simultaneously detecting four tumor markers and preparation method of chemiluminescent kit

Abstract

The invention relates to the technical field of immune detection and provides a chemiluminescent kit for simultaneously detecting four tumor markers and a preparation method of the chemiluminescent kit. The preparation method comprises the following steps: preparing an antibody coating; respectively pouring, sealing, washing and airing an enzyme-linked immunosorbent assay (ELISA) plate in the first step, and storing the ELISA plate for later use; charging a sample, putting the ELISA plate into an incubation oscillator, and incubating an antibody; adding into kit streptavidin-horse radish peroxidase (SA-HRP); shooting an image through a charge coupled device (CCD) imager, and calculating an IDV value of the image through image analysis software; drawing a standard curve according to the IDV value, and calculating a sample concentration result; and performing calculation and statistical analysis on the sample concentration result by using a softMax regression model and an Excel form, and performing comprehensive analysis. By virtue of the chemiluminescent kit and the preparation method thereof, the problems of complicated process, inconvenience in operation and low sensitivity are effectively solved.

Description

technical field [0001] The invention relates to the technical field of immunoassay, in particular to a chemiluminescent reagent kit for simultaneously detecting four tumor markers and a preparation method thereof. Background technique [0002] In recent years, more and more evidences have proved the role of inflammatory response in the occurrence and development of depression. Many inflammatory factors and corresponding soluble receptors, chemotactic molecules, adhesion molecules, acute phase proteins, etc. Peripheral blood and cerebrospinal fluid in patients with depression significantly increased, and the relationship between individual symptoms of depression, such as fatigue, cognitive impairment, and sleep disturbance, and inflammatory factors has also been continuously reported. These depression-related biomarkers can be used to assist clinical diagnosis and prognosis evaluation, and will provide biological evidence for the objective diagnosis of depression. Further, w...

AI Technical Summary

Problems solved by technology

However, in the usual enzyme-linked immunosorbent assay (ELISA) detection of samples, each detection well generally requires 100-200 μl, and the sample volume is relatively large. In addition, there are more than one biomarkers of depression, which requires joint monitoring of multiple factors for diagnosis. If you still rely on a single ELISA test, the sample size will be too large; it is obviously time-consuming and laborious to test each factor individually to assist the diagnosis of depression, and the cost of diagnosis is high, which is generally unaffordable for patients

[0004] RIA technology, the radioimmunoassay method, has both high specificity of immune response and high sensitivity of radioactivity measurement, but there are problems such as radiation radiation and pollution, and it also poses a threat to the health of testing personnel, and the operation is relative to ELISA Complicated, only one indicator can be measured in one inspection, and a special radioactive experiment site needs to be provided, which requires high investment and great harm

[0005] Although the protein chip can realize the function of simultaneous detection of multiple factors, it is because it coats the antibody on the substrate slide, microsphere or membrane used to prepare the protein chip, and the substrate slide and membrane must be aldehyde-treated before use. Kylation or poly-lysine and other pretreatments lead to increased production costs, and the follow-up experiments are cumbersome, poor repeatability, and difficult to promote

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