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A method for constructing a probe for detection of Alzheimer's disease marker Aβ oligomer by colorimetric method based on nucleic acid aptamer

An Alzheimer's disease, nucleic acid aptamer technology, applied in the field of biological nanometers, can solve the problems of insufficient timeliness, lack of molecular platform, and reduced sensitivity of electrochemical methods, and achieves low detection limit, low cost, and detection. low limit effect

Active Publication Date: 2019-04-05
CENT SOUTH UNIV
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

The existing electrochemical, fluorescence, and SPR detection methods for Aβ all have their advantages and disadvantages. Effectiveness, ease of operation, and no radiation are common advantages. However, since Aβ species are usually Aβ monomers and Aβ oligomers, A mixture of Aβ aggregates, if the detection method cannot accurately identify Aβ oligomers, it will reduce its sensitivity and is not time-sensitive enough for electrochemical methods
At the same time, there are currently methods to detect Aβ oligomers by computer technology, but due to the lack of molecular platforms, it is still a great challenge to design a molecule to detect Aβ oligomers in Aβ species

Method used

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  • A method for constructing a probe for detection of Alzheimer's disease marker Aβ oligomer by colorimetric method based on nucleic acid aptamer
  • A method for constructing a probe for detection of Alzheimer's disease marker Aβ oligomer by colorimetric method based on nucleic acid aptamer
  • A method for constructing a probe for detection of Alzheimer's disease marker Aβ oligomer by colorimetric method based on nucleic acid aptamer

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Embodiment 1

[0032] (1) Preparation of gold nanoparticles and the combination of gold nanoparticles and aptamers to prepare probes, the steps are as follows:

[0033] First, heat 100mL of 0.01% chloroauric acid to boiling under stirring, then quickly add 2mL of 2% sodium citrate solution to the boiling solution, and continue to stir to make it evenly mixed. At this time, the stirring should not be too vigorous to avoid uneven mixing , and keep boiling at the same time, when the color changes from colorless to wine red, start timing, continue heating for 15 minutes, then take the solution out of the oil bath, place it on a magnetic stirrer and stir to cool to room temperature, and then use ultraviolet spectroscopy A photometer and a particle size analyzer were used to characterize the particle size and degree of dispersion of the nano-gold solution.

[0034] (2) Binding of gold nanoparticles and aptamers

[0035] The Alzheimer's disease marker aptamer is reacted with gold nanoparticles at ...

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Abstract

The invention discloses a method for detecting an Alzheimer's disease marker by adopting a colorimetric method based on a nucleic acid aptamer. The method comprises the steps that the Alzheimer's disease marker aptamer is modified on the surface of nano gold to obtain a probe; a series of Abeta oligomer standard solutions different in concentration respectively react with the probe and then respectively react with a NaCl solution, ultraviolet absorption peak values of an aggregation state and a dispersing state of the nano gold of the reaction liquid are detected through ultraviolet, a standard curve about Abeta oligomer concentration values and the corresponding ratio of the ultraviolet absorption peak value of the aggregation state of the nano gold to the ultraviolet absorption peak value of the dispersing state of the nano gold is established, the solution to be detected is detected, analysis is performed based on the standard curve, and then the concentration of Abeta oligomers in the solution to be detected is obtained. The method can achieve concentration detection of the Alzheimer's disease marker and has the advantages of being good in stability, low in cost and fast.

Description

technical field [0001] The invention relates to a method for detecting Alzheimer's disease markers, in particular to a method for detecting Alzheimer's disease markers based on a nucleic acid aptamer colorimetric method; it belongs to the field of biological nanometers. Background technique [0002] Alzheimer's disease has become one of the major diseases threatening human health in the 21st century. It is a degenerative disease characterized by gradual cognitive decline, irreversible harmful impairment and observable memory impairment. There are senile plaques deposited in the brain of Alzheimer's patients, and Aβ amyloid is the main component of senile plaques. Therefore, the degree of senile dementia can be estimated by detecting Aβ amyloid. Synaptic toxicity, and levels in the cerebrospinal fluid of Alzheimer's patients are significantly higher than normal. Therefore, it is particularly important to develop a method that can quickly detect the content of Aβ oligomers, h...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/33G01N33/68
CPCG01N21/33G01N33/6896G01N2800/2821
Inventor 邓春艳刘慧向娟钱频
Owner CENT SOUTH UNIV
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