Electrochemical luminescence sensor system for determining adenosine triphosphate and preparation method and application thereof

An adenosine triphosphate, electrochemical technology, applied in the field of biomedical detection, can solve the problems of high cost, tedious time-consuming, low sensitivity, etc.

Active Publication Date: 2020-04-17
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Purpose of the invention: In view of the shortcomings of the prior art, such as cumbersome time-consuming, low sensitivity, bulky equipment, and high cost, the present invention provides an electrochemiluminescent sensor system for the determination of adenosine triphosphate. Electrochemiluminescent sensor system with multiple signal amplification strategies

Method used

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  • Electrochemical luminescence sensor system for determining adenosine triphosphate and preparation method and application thereof
  • Electrochemical luminescence sensor system for determining adenosine triphosphate and preparation method and application thereof
  • Electrochemical luminescence sensor system for determining adenosine triphosphate and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0094] Au@Fe 3 o 4 - Preparation of substrate-aptazyme probes

[0095] (1) Fe 3 o 4 Preparation of nanoparticles

[0096] Weigh 0.65g FeCl 3 and 0.2g of trisodium citrate into a beaker filled with 20mL of ethylene glycol, stir well until it is completely dissolved, add 1.2g of sodium acetate into it, carry out magnetic stirring for 30min, add the above mixture into the reaction kettle, and react at 200°C 10h; the precipitate was washed with ethanol and water three times successively after magnetic separation to obtain Fe 3 o 4 Nanoparticles.

[0097] (2) Preparation of Au nanoparticles

[0098] Configure 50mL mass fraction as 0.01% HAuCl 4 Add the aqueous solution into a round-bottomed flask, heat it to boiling, add 1 mL of 0.1% trisodium citrate aqueous solution to the solution, continue heating and boiling for 15 min, and cool the sample to room temperature to obtain an Au nanoparticle solution.

[0099] (3)Au@Fe 3 o 4 Nanoparticle preparation

[0100] Weigh 10m...

Embodiment 2

[0104] RuSiO 2 -CS solution was prepared as follows:

[0105] Measure 2mL Triton X-100, 8mL cyclohexane, 2mL n-hexanol into the reaction vessel, mix well, add 350μL of 40mM Ru(bpy) to the mixture 3 Cl 2 Aqueous solution, after mixing, add 150μL TEOS and 100μL ammonia water, stir and react for 24h; add 5mL acetone, centrifuge, wash the precipitate with ethanol and water in turn, and resuspend the product in ethanol to obtain 4mg / mL RuSiO 2 solution.

[0106] Add 1mg CS to 1mL 2% acetic acid aqueous solution, ultrasonically disperse for 20min to obtain CS solution; take 1mL 4mg / mL RuSiO 2 Aqueous solution was added to 1mL CS solution, sonicated for 30min to obtain uniformly dispersed RuSiO 2 -CS solution.

Embodiment 3

[0108] Preparation method of electrochemiluminescent sensor for measuring Trigger DNA

[0109] The preparation method of electrochemiluminescent sensor is as follows: figure 1 shown, including the following steps:

[0110] (1) Electrode pretreatment: use 0.3μm and 0.05μm Al in turn for the working electrode 2 o 3 After the powder is polished, ultrasonically clean it with water, ethanol, and water for 4 minutes;

[0111] (2) Modified RuSiO 2 : Take 10μL 2mg / mL RuSiO 2 - CS solution (Example 2) was added dropwise to the surface of the above-mentioned working electrode, left to dry at room temperature, washed with PBS solution, and dried in the air;

[0112] (3) Covalently link Capture DNA: dropwise add 2.5% glutaraldehyde aqueous solution to the surface of the above electrode, react at room temperature for 2 hours, wash with PBS solution, and dry in the air; add 10 μL of 4 μM Capture DNA dropwise to the electrode surface, 37 React at ℃ for 2 hours, wash with PBS solution, ...

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Abstract

The invention discloses an electrochemical luminescence sensor system for determining adenosine triphosphate and a preparation method and application thereof. The system is composed of a magnetic probe for amplifying an adenosine triphosphate concentration signal and an electrochemical luminescence sensor for determining Triger DNA. The magnetic probe is formed by compounding Fe3O4 nano particles,gold nano particles, a DNA substate and aptazyme in sequence. The electrochemical luminescence sensor is obtained by connecting amino-modified Capture DNA to the surface of a RuSiO2-CS-modified working electrode through glutaraldehyde cross-linking action. According to the invention, adenosine triphosphate participates in an isothermal amplification reaction guided by the magnetic probe and a large amount of intermediate Trigger DNAs are generated; and then, the intermediate Triger DNAs are quantitatively detected through the electrochemical luminescence sensor; and finally, the purpose of indirectly and quantitatively detecting adenosine triphosphate is achieved.

Description

technical field [0001] The invention belongs to the field of biomedical detection, and in particular relates to an electrochemiluminescent sensor system for measuring adenosine triphosphate, a preparation method and application thereof. Background technique [0002] Adenosine triphosphate (ATP) is a small molecular high-energy phosphate compound, and it is also the main energy source in organic life, ensuring the energy supply for various life activities of cells. As an "energy currency", it participates in many important physiological processes in living organisms, such as: gene synthesis, nutrient metabolism, drug delivery, and regulation of biological activities such as immunity and nerve mediation. Relevant studies have shown that ATP is an indicator of cell viability and cell damage in organic organisms, and changes in its content are closely related to the occurrence of cardiovascular diseases, Parkinson's disease, Alzheimer's disease and other diseases. In addition, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6825C12Q1/682G01N27/30G01N21/76G01N27/416
CPCC12Q1/6825C12Q1/682G01N27/30G01N21/76G01N27/4163C12Q2565/607C12Q2563/137C12Q2563/155C12Q2531/119C12Q2563/103
Inventor 沈艳飞宁振强
Owner SOUTHEAST UNIV
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