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A kind of aptamer sensor based on charge effect and its construction method and application

An aptamer sensor and construction method technology, which is applied in the direction of instruments, scientific instruments, and electrochemical variables of materials, can solve the problems of high cost, reduced sensitivity, and easy fall-off of modified sulfhydryl molecules, and achieve improved sensitivity, low detection limit, The effect of high sensitivity

Inactive Publication Date: 2019-07-30
CENT SOUTH UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The existing fluorescence and SPR methods for detecting Aβ have their advantages and disadvantages. Effectiveness, ease of operation, and no radiation are common advantages. However, Aβ species are usually Aβ monomers, Aβ oligomers, and Aβ aggregates. If the detection method cannot accurately identify Aβ oligomers, it will reduce its sensitivity
For the traditional electrochemical method, the typical method is to use mercapto groups to combine with gold electrodes and fix them on the electrode surface. This method is unstable and easy to fall off during electrochemical testing, and the molecular cost of modifying mercapto groups is relatively high.
Therefore, rapid, sensitive, and low-cost detection of Aβ oligomers remains a great challenge

Method used

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  • A kind of aptamer sensor based on charge effect and its construction method and application
  • A kind of aptamer sensor based on charge effect and its construction method and application
  • A kind of aptamer sensor based on charge effect and its construction method and application

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Effect test

Embodiment 1

[0043] (1) The method for the biosensor constructed on the electrode, the steps are as follows:

[0044]Take a small centrifuge tube, mix 5 μL of 50 μM 4-[(trimethylsilyl)ethynyl]benzoic acid THF solution with 50 μL of 20 μM tetrabutylammonium fluoride THF solution, then invert on the polished electrode, Soak the electrode completely in the mixed solution and react for 1h at room temperature to form Au-C bonds.

[0045] Prepare 10 μM DNA1 and 10 μM DNA2 solutions respectively, and mix and incubate them at 37°C for 2 hours to obtain a dsDNA mixture.

[0046] The electrode was rinsed with THF, and the incubated dsDNA solution was mixed with 50 μM EDC (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride) and 50 μM NHS (N-hydroxysuccinate imide) solution mixed with the same volume, put it upside down on the electrode, and let it stand for 3 hours. The amide reaction between the amino group on DNA1 and the -COOH of 4-[(trimethylsilyl)ethynyl]benzoic acid occurs, thereby ma...

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Abstract

The invention discloses a charge effect-based aptamer sensor, and a construction method and an application thereof. The construction method of the sensor comprises the following steps: immersing a gold electrode in a tetrahydrofuran solution containing 4-[(trimethylsilyl)ethynyl]benzoic acid and tetrabutylammonium fluoride, and carrying out a reaction in a room temperature environment to obtain a 4-carboxylphenylethynyl modified gold electrode; carrying out a hybridization reaction on DNA1 and DNA2 to obtain dsDNA; and carrying out an amidation reaction on the 4-carboxylphenylethynyl modified gold electrode and the dsDNA, and adding a compound containing nanogold-streptavidin, ferrocenylalkanethiol and a tumor marker aptamer to obtain the aptamer sensor having the advantages of low cost, good stability, simplicity in detection, high sensitivity and high specificity.

Description

technical field [0001] The invention relates to a biosensor, in particular to an aptamer sensor based on a charge effect and a construction method thereof, and an electrochemical method for detecting tumor markers by using the sensor; it belongs to the field of electrochemical biosensors. Background technique [0002] Alzheimer's disease (AD) is a progressive and fatal neuromutative disease. However, there is no definitive diagnostic test in routine clinical practice. Markers of AD brain include plaques composed of extracellular β-amyloid (Aβ) protein aggregates and intracellular hyperphosphorylated tau neurofibrillary tangles. The current research on the amyloid Aβ affecting AD shows that the aggregation process of Aβ is roughly the aggregation process of Aβ from monomer to oligomer and then to fiber. Compared with amyloid fibrils, soluble amyloid oligomers are the most cytotoxic aggregates and are significantly higher in the CSF of Alzheimer's patients than in normal ind...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/327G01N27/48
CPCG01N27/3277G01N27/3278G01N27/48
Inventor 向娟刘慧邓春艳
Owner CENT SOUTH UNIV
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