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Signal-amplification fluorescence detection system, fluorescence biosensor and application thereof

A fluorescence detection and signal amplification technology, applied in fluorescence/phosphorescence, instruments, measuring devices, etc., can solve the problems of limited quantitative accuracy, cumbersome detection steps, and high detection cost, and achieve improved accuracy, high signal-to-noise ratio, and high The effect of detection sensitivity

Pending Publication Date: 2020-08-11
SHENZHEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] Therefore, the technical problem to be solved by the present invention is to overcome the defects of high detection cost, cumbersome detection steps, and limited quantitative accuracy in the disease diagnosis of Alzheimer's disease in the prior art.

Method used

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  • Signal-amplification fluorescence detection system, fluorescence biosensor and application thereof
  • Signal-amplification fluorescence detection system, fluorescence biosensor and application thereof
  • Signal-amplification fluorescence detection system, fluorescence biosensor and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] This embodiment provides a fluorescence detection system, including the following components:

[0045] 1. The recognition unit is a single-stranded DNA probe labeled with a fluorescent molecule. Specifically, the recognition unit includes: a streptavidin-labeled fluorescent molecule, a biotin-labeled single-stranded DNA molecule (ssDNA), and streptomycin is coupled with biotin to form a recognition unit. Wherein, the fluorescent molecules are specifically fluorescent quantum dots (CdSe / ZnS quantum dots), and the coupling molar ratio of streptavidin-labeled fluorescent molecules to biotin-labeled single-stranded DNA molecules is 1:4. The nucleotide sequence of the single-stranded DNA molecule is selected from any nucleotide sequence shown in SEQ ID NO.1-SEQ ID NO.3, specifically selects the nucleotide sequence of SEQ ID NO.1 in this embodiment, has The recognition unit of the nucleotide sequence of SEQ ID NO.1 can recognize and bind to the biomarker molecule miRNA1 of A...

Embodiment 2

[0052] This embodiment provides a fluorescent biosensor, including the fluorescence detection system in Embodiment 1. Fluorescent biosensors can realize the rapid detection of biomarker molecules of Alzheimer's disease, and have the advantages of fast, simple, sensitive, efficient and accurate, and can be used for disease screening of Alzheimer's disease with diagnosis.

experiment example 1

[0054] 1. Experimental purpose: To optimize the conditions of the fluorescence detection system in Example 1 to improve the detection performance.

[0055] 2. Experimental process and results:

[0056] Fluorescence detection was performed using a multifunctional microplate reader Variokan Flash (Thermo Scientific, USA) in a black 96-well plate. 100 μL of different concentrations of miRNA (1000 nM, 500 nM, 200 nM, 100 nM, 50 nM, 20 nM, 10 nM and OnM) were added to the fluorescence detection system and incubated for 10 minutes. Then add DnaseI to each well and incubate for 30 minutes. During the detection process, the fluorescence emission intensity at 520 nm (excitation wavelength: 380 nm) was recorded every five minutes. Change the addition amount of graphene oxide (GO) and DNase I in the fluorescence detection system, compare three concentrations of GO (500μg / ml, 200μg / ml, 100μg / ml) and five kinds of DNase I additions (10U, The fluorescence detection results under the cond...

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Abstract

The invention discloses a signal-amplification fluorescence detection system which comprises a recognition unit, which is marked with fluorescent molecules, is used for recognizing biomarker moleculesof Alzheimer's disease, and is combined with the biomarker molecules to form a biological complex; a fluorescence quenching unit which is used for adsorbing the recognition unit to quench fluorescence of the fluorescent molecules, wherein the fluorescence quenching unit and the biological complex are desorbed, so that fluorescence of fluorescence molecules is recovered; and a signal amplificationunit, which is used for degrading the recognition unit in the biological complex. The fluorescence detection system is used for detecting the biomarker molecules of the Alzheimer's disease, the fluorescence detection system has the advantages of rapidness, simplicity, sensitivity and high efficiency; and meanwhile, the system can amplify a fluorescent molecular signal and reduce the detection limit of the system, and has high sensitivity. The invention further discloses a fluorescence biosensor which comprises the fluorescence detection system and can provide effective information for diagnosis and screening of the Alzheimer's disease.

Description

technical field [0001] The invention relates to the field of biomolecular detection, in particular to a fluorescent detection system for signal amplification, a fluorescent biosensor and applications thereof. Background technique [0002] Alzheimer's disease (AD) is an irreversible neurodegenerative disease mainly related to age. Alzheimer's disease affects the patient's cognition and short-term memory function. As the disease progresses, the patient's daily life ability will be gradually affected, and eventually the patient will lose the ability to take care of himself. In terms of epidemiology, the incidence of Alzheimer's disease increases exponentially with age. With the increasing aging trend in the world, Alzheimer's disease has become the most serious disease and cause of death after tumor and cardiovascular disease. [0003] Due to the long incubation period of Alzheimer's disease, which can reach about 10-20 years, many patients miss the precious opportunity for e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6402G01N21/6428
Inventor 周洁许改霞
Owner SHENZHEN UNIV
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