Labeling method of converting recognition signal of biomarker into visible light color spectrum, and application

A biomarker and signal recognition technology, applied in clinical laboratory medicine, rapid detection, and biomedical fields, can solve the problems of unfavorable primary hospitals and family rapid diagnosis methods, increase the difficulty of experimental operation, and the difficulty of waste disposal, etc., to achieve the benefit of Visual recognition, low cost, and the effect of signal recognition

Pending Publication Date: 2019-01-22
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] (1) Radioimmunolabeling technology: there is weak radioactive contamination, and waste disposal is difficult; the existence of the half-life of radioisotopes increases the difficulty of experimental operation;
[0005] (2) ELISA labeling technology: enzyme markers are easy to inactivate; the sensitivity is not high; the reaction process is relatively complicated;
[0006] (3) Fluorescent immunolabeling technology: the reaction process is rapid, but at the same time it is easily affected by light scattering and endogenous fluorescent substances in the sample;
[0007] (4) Luminescent immunolabeling technology: additional solid-phase carriers are required, and the operation process is relatively complicated;
[0008] (5) Colloidal gold immunolabeling technology: the sensitivity is affected, the technology must be high, and the operation process is difficult
[0009] To sum up, in view of the unfavorable rapid diagnosis method of grass-roots hospitals and families caused by labeling, this invention designs a new labeling method based on immune analysis, which can quickly diagnose, Pre-bed diagnosis (POCT), especially important for children and the elderly

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] 10,12-Pentacosadiynoic acid (PCDA) was dissolved in absolute ethanol, and liposomes were prepared by thin-film ultrasonic method. The solvent in the prepared liposome solution was PBS buffer (10 mM), and the liposome concentration was 1 mM. Store overnight at 4°C in the dark, and obtain blue polymer liposomes by ultraviolet crosslinking. The polyclonal antibody that can specifically recognize the β-isomeric CTX fragment was dissolved in PBS buffer at a concentration of 10 mM, mixed with the liposome solution, and carbodiimide (EDC) and N-hydroxysuccinimide (NHS) were added. ) (concentration ratio EDC:NHS:liposome=2:1:1), the probe was coupled to the liposome surface, centrifuged to remove free antibody in the solution, and stored in the dark.

[0052] Add 6 μl of 1mg / ml sample containing target detection substance, incubate at 25°C for 30 minutes, the color changes from blue to red.

Embodiment 2

[0054] 10,12-nonacosadiynoic acid (10,12-nonacosadiynoic acid) was dissolved in chloroform and mixed with phospholipid molecules, and liposomes were prepared by injection method. The solvent in the prepared liposome solution was PBS buffer (10 mM), and the liposome concentration was 0.5 mM. Store overnight at 4°C in the dark, and obtain blue polymer liposomes by ultraviolet crosslinking. The polyclonal antibody that can specifically recognize the CTX fragment is dissolved in the PBS buffer solution with a concentration of 10 mM, mixed with the liposome solution, and glutaraldehyde with a mass fraction of 25% is added to couple the probe to the surface of the liposome. Centrifuge to remove free antibody in the solution, and store in the dark.

[0055] Add 4 μl of 1mg / ml sample containing target detection substance, incubate at 25°C for 30 minutes, the color changes from white to red.

Embodiment 3

[0057] 10,12-Tricosadiynoic acid (10,12-Tricosadiynoic acid) was dissolved in chloroform, and liposomes were prepared by thin-film ultrasonic method. The solvent in the prepared liposome solution was PBS buffer (10 mM), and the liposome concentration was 0.5 mM. Store overnight at 4°C in the dark, and obtain blue polymer liposomes by ultraviolet crosslinking. Dissolve the polyclonal antibody that can specifically recognize type I collagen in Tris buffer with a concentration of 10 mM, mix it with the liposome solution, add glutaraldehyde with a mass fraction of 25%, and couple the probe to the liposome surface, centrifuge to remove free antibody in the solution, and store in the dark.

[0058] Add 6 μl of 0.5mg / ml sample containing target detection substance, incubate at 25°C for 30 minutes, the color changes from gray to purple.

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PUM

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Abstract

The invention discloses a labeling method of converting a recognition signal of a biomarker into visible light color spectrum, and application. Liposome with a force-induced discoloration characteristic is prepared from diacetylene monomer and phospholipid molecules, the liposome modifies the surface of a probe molecule capable of recognizing an antibody, agglutinant or peptide molecules of to-be-detected marker specifically, the probe molecule and a target to-be-detected marker react, so as to cause the liposome to undergo force-induced discoloration to enable the probe molecule to convert arecognition signal of a to-be-detected biomarker into visible light color spectrum that can be seen by naked eyes, and qualitative determining is performed according to color change of the solution. The marker changes greatly in color in a range of visible light, fewer components are used, the cost is lower, and reaction is faster.

Description

technical field [0001] The invention belongs to the technical fields of biomedicine, clinical laboratory medicine and rapid detection, and relates to a marking method and application for converting identification signals of biomarkers into visible light chromatograms. Background technique [0002] With the advent of an aging society, orthopedic diseases are increasingly harmful to the human body, and the requirements for early diagnosis and process control of related diseases are also getting higher and higher. Pathologically, we have a clear understanding of human bone metabolism and related diseases, and comprehensive assessment of the physiological state of bone (bone formation and resorption) is the basis for judging bone development, bone attenuation and bone loss, and thus for the diagnosis and analysis of bone diseases. Among them, type I collagen is one of the most important components in bone structure. During bone matrix metabolism in bone tissue, type I collagen i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/78G01N33/533G01N33/577
CPCG01N21/78G01N33/533G01N33/577
Inventor 蒋波邓啸鸣冯欢欢李晓磊李霞
Owner SICHUAN UNIV
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