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Detection method for core-shell quantum dot-based flexible coupled marker immunochromatography test strip

An immunochromatographic test paper and core-shell quantum dot technology, applied in the field of immunoassay, can solve the problems of unsatisfactory results in quantitative detection sensitivity and specificity, weakened fluorescence characteristics, and reduced detection effect, etc., and achieves strong photochemical stability and emission. Narrow spectral lines and high fluorescence quantum efficiency

Inactive Publication Date: 2014-04-30
CHANGCHUN INST OF OPTICS FINE MECHANICS & PHYSICS CHINESE ACAD OF SCI
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Problems solved by technology

However, quantum dots are a kind of nanomaterials with great surface effects. Existing test strips using quantum dots are biomarked by quantum dots and antibody molecules through electrostatic adsorption and direct covalent connection. In the process of labeling the antibody, it is easy to label the Fab end of the antibody with the epitope site. The labeled antibody cannot immunoreact with the antigen molecule, so it is an invalid label. This direct connection technology is very important for the final The sensitivity of immunoassay, especially the specificity, will be greatly affected, reducing the detection effect, and the sensitivity and specificity of quantitative detection cannot achieve satisfactory results
Moreover, the existing quantum dot test strip patents all use bare-core quantum dots for immunoassay. Since the fluorescence characteristics of bare-core quantum dots will be greatly weakened compared with core-shell quantum dots, it is applied to test strip immunoassays. detection, the effect must be weaker than the results of immunoassay with core-shell quantum dots

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  • Detection method for core-shell quantum dot-based flexible coupled marker immunochromatography test strip
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[0025] The preparation method of the core-shell quantum dots described in the first step of the present invention is a prior art, and the method utilizes continuous ion layer adsorption reaction technology (SILAR) to realize the core-shell coating of the quantum dots. Adopt the principle of one-pot method, add nuclear raw materials into the same reaction device, obtain bare-nuclear quantum dots with different particle sizes by controlling the time of nuclear growth, then add shell raw materials drop by drop, use SILAR technology for shell coating, and finally The samples were centrifuged and purified to obtain core-shell quantum dots. For specific methods, refer to the article J.Phys.Chem.C2008, 112, 8587-8593.

[0026] The core-shell quantum dots described in the present invention are preferably core-shell quantum dots such as CdSe / CdS, CdSe / ZnSe, and CdSe / ZnS. By changing the particle size of the quantum dots, the tuning of the fluorescent color can be realized, and it is mor...

Embodiment 1

[0038] 1. Weigh 0.051g (0.4mmol) of tellurium powder and 0.08g (2mmol) of sodium borohydride, and add them into 10ml of distilled water which is fully deoxygenated with nitrogen to obtain a solution of sodium tellurium hydride;

[0039] 2. Weigh 0.6714 g (1.6 mmol) of cadmium perchlorate hexahydrate, add it to a three-necked bottle containing 100 ml of anaerobic water, then add 342 microliters (3.84 mmol) of mercaptopropionic acid, and use 1 mol / L sodium hydroxide The pH value of the solution is adjusted between 10-11;

[0040] 3. Take the sodium telluride hydride solution synthesized in step 1 and inject it into the three-necked bottle in step 2, raise the temperature to 100°C, synthesize CdTe bare-nucleus quantum dots, and obtain naked nuclei within 15-30 minutes by changing the reaction time A green solution containing green light quantum dots with a size less than 3nm;

[0041] 4. Repeat step 2 to prepare the shell material of cadmium; dissolve the sodium sulfide nonahydr...

Embodiment 2

[0048] 1. Weigh 0.051g (0.4mmol) of selenium powder and 0.08g (2mmol) of sodium borohydride, and add them into 10ml of distilled water that is fully deoxygenated with nitrogen to obtain a solution of sodium telluride hydride;

[0049] 2. Weigh 0.6714 g (1.6 mmol) of cadmium perchlorate hexahydrate, add it to a three-necked bottle containing 100 ml of anaerobic water, then add 342 microliters (3.84 mmol) of mercaptopropionic acid, and use 1 mol / L sodium hydroxide The pH value of the solution is adjusted between 10-11;

[0050] 3. Take the sodium selenium hydride solution synthesized in step 1 and inject it into the three-necked bottle in step 2, raise the temperature to 100°C, synthesize CdSe bare-nucleus quantum dots, and obtain naked nuclei within 15-30 minutes by changing the reaction time A green solution containing green light quantum dots with a size less than 3nm;

[0051] 4. Repeat step 2 to prepare the shell material of cadmium; dissolve the sodium sulfide nonahydrate...

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Abstract

The invention provides a detection method for a core-shell quantum dot-based flexible coupled marker immunochromatography test strip, belonging to the technical field of an immunodetection method. According to the method, based on the fact that the core-shell quantum dot has the characteristics of being wide in exciting line, narrow in emission spectrum line, high in quantum size effect and fluorescent quantum efficiency and strong in photochemical stability, quantum dots and protein G or protein A molecules are subjected to covalent linkage by adopting an EDC condensation mode, the flexible coupling of the quantum dots and antibody molecules can be realized by utilizing protein G or protein A molecules as bridges and by means of the specificity of Fc terminal of an antibody, antigenic determinant part of the antibody is not occupied, the antigen immunity activity of the antibody can be greatly improved, and the sensitivity and specificity when the marker product is utilized for immunodetection can be greatly enhanced.

Description

technical field [0001] The invention belongs to the technical field of immunodetection methods, in particular to a detection method based on core-shell quantum dot flexible coupling marker immunochromatographic test strips. Background technique [0002] At present, the detection test strips using nano-gold color development have been widely used. The basic principle is that the microporous membrane is used as the solid phase carrier, and the antigen or antibody of the detection object is coated. After adding the sample to be tested, it passes through the capillary of the microporous membrane. The siphon action makes the detection substance in the sample combine with the antigen or antibody coated on the membrane, and the detection result is judged by the color of the gold nanometer marker itself. However, for some samples with low content, the color of gold nanoparticles is very light, it is difficult to judge the results with the naked eye, and the sensitivity is low, so it...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/543G01N33/558G01N33/588
Inventor 曾庆辉纪文宇
Owner CHANGCHUN INST OF OPTICS FINE MECHANICS & PHYSICS CHINESE ACAD OF SCI
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