Method for fluorescence detection of Alzheimer's disease markers based on silver nano-cluster probe containing repeated AGGGTT sequences

A silver nanocluster and fluorescence detection technology, which is applied in fluorescence/phosphorescence, material excitation analysis, etc., can solve the problems of sensitivity and detection concentration range to be improved

Inactive Publication Date: 2017-12-05
CENT SOUTH UNIV
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Problems solved by technology

Monomers, oligomers, and fibrils of β-amyloid protein are the main markers of Alzheimer's disease and widely exist in cerebrospinal fluid. It has a posi

Method used

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  • Method for fluorescence detection of Alzheimer's disease markers based on silver nano-cluster probe containing repeated AGGGTT sequences
  • Method for fluorescence detection of Alzheimer's disease markers based on silver nano-cluster probe containing repeated AGGGTT sequences
  • Method for fluorescence detection of Alzheimer's disease markers based on silver nano-cluster probe containing repeated AGGGTT sequences

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Embodiment 1

[0054] dna 1 -AgNCs preparation method, the steps are as follows:

[0055] Take 15 μL of DNA 1 (100μM) mixed with 73uL phosphate buffer (20mM, pH 6.5), add 6μL AgNO 3 Shake and mix the aqueous solution (1.5mM) at room temperature for 15min, then add 6μL freshly prepared NaBH 4 (1.5mM) mixed quickly and transferred to a constant temperature water bath at 25°C to react for 2h, then its fluorescence can be detected.

[0056] dna 1 / DNA 2 -AgNCs reaction steps are as follows:

[0057] Take 5 μL DNA 2 (100μM) with 50μL DNA 1 -AgNCs solution was mixed and reacted in a constant temperature water bath at 25°C for 1 hour to detect its fluorescence signal.

[0058] dna 1 / DNA 2 -AgNCs-Cu 2+ Probe preparation steps are as follows:

[0059] Add 6uL copper nitrate (0.156mM) to the reacted DNA 1 / DNA 2 -AgNCs solution (55 μL) was mixed and reacted in a constant temperature water bath at 25° C. for 0.5 h to detect its fluorescence signal.

[0060] Aβ 1-40 The steps to establi...

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Abstract

The invention discloses a method for the fluorescence detection of Alzheimer's disease markers based on a silver nano-cluster probe containing repeated AGGGTT sequences. The method comprises the following steps: synthesizing a silver nanocluster by using DNA1 as a template, specifically binding the DNA1 with DNA2, binding the bound DNA1 and DNA2 with copper ions to obtain a DNA1/DNA2-AgNCs-Cu<2+> probe, and using the DNA1/DNA2-AgNCs-<2+> fluorescence probe as a probe for detecting the Alzheimer's disease markers to detect the content of the Alzheimer's disease markers Abeta 1-40 monomers and Abeta1-40 oligomers through the change of fluorescence signals. The method realizes the fluorescence detection of the Alzheimer's disease markers, and has the advantages of stable signal, strong resistance to environmental impact, high sensitivity, low detection limit, good linear relationship, and easiness in operation.

Description

technical field [0001] The present invention relates to a method for detecting Alzheimer's disease markers, in particular to a method for constructing a silver nanocluster probe containing a repeating AGGGTT sequence, and using the silver nanocluster probe containing a repeating AGGGTT sequence to detect Alzheimer's disease. A method for fluorescent detection of Alzheimer's disease markers, which uses Alzheimer's disease markers and DNA 1 / DNA 2 -AgNCs-Cu 2+ The principle of combining copper ions in the probe and changing the fluorescent signal to detect the content of Alzheimer's disease markers belongs to the field of bionano. technical background [0002] Alzheimer's disease has become one of the major diseases threatening human health in the 21st century, and the successful detection of low levels of Alzheimer's disease markers has become an urgent problem to be solved. Monomers, oligomers, and fibrils of β-amyloid protein are the main markers of Alzheimer's disease a...

Claims

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Application Information

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IPC IPC(8): G01N21/64
Inventor 向娟张秀姣邓春艳
Owner CENT SOUTH UNIV
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