Adenosine detecting method based on micro-fluidic chip and nucleic acid adapter technology

A technology of microfluidic chip and nucleic acid aptamer, which is applied in the field of biological detection, can solve the problems of limited application and limited detection sensitivity in the field of high-sensitivity microanalysis, and achieve the effect of specific and accurate quantitative analysis.

Inactive Publication Date: 2014-04-09
HUNAN INSTITUTE OF ENGINEERING
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Problems solved by technology

Traditional adenosine detection methods mainly include HPLC, ultraviolet detection, flow cytometry and fluorescence analysis. These technologies have l...

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  • Adenosine detecting method based on micro-fluidic chip and nucleic acid adapter technology
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  • Adenosine detecting method based on micro-fluidic chip and nucleic acid adapter technology

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Embodiment Construction

[0022] The technical solutions of the present invention will be further described below in conjunction with the accompanying drawings and specific embodiments.

[0023] refer to figure 1 , is a schematic diagram of the principle of the method of the present invention. figure 1 The specific principles shown are as follows:

[0024] First, biotin-modified nucleic acid aptamer 3 and biotin-modified casein 2 are modified to the surface of avidin-modified microspheres 1 according to a certain ratio, and functionalized microspheres 4 that can be used for adenosine detection are prepared; influx modification Functional nanoparticles containing horseradish peroxidase (HRP) and capture probes 5 are hybridized with the functionalized microspheres in the chip, and the capture probes and nucleic acid aptamers are combined by hybridization to immobilize the functionalized nanoparticles on the chip. Microsphere surface 6; flow into the target molecule adenosine 7, adenosine and nucleic ac...

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Abstract

The invention discloses an adenosine detecting method based on a micro-fluidic chip and a nucleic adapter technology. Biotin-modified adenosine nucleotide adapter and biotin-modified casein are fixed on the surface of an avidin-modified microsphere to form a functional microsphere through a biotin-avidin combining method; capture probes modified with horseradish peroxidase and sulfydryl are modified on nanometer particles as signal marks; functional nanometer particles flow to a micro-fluidic microsphere array to detect a detection area of a chip and hybrid with a microsphere array; after elution, an adenosine solution flows to be incubated with hybridized microspheres; a peroxidase substrate solution flows in; biotinylated tyramine is combined with the casein on the surface of the microsphere; the avidin-marked quantum dot flows in after elution; qualification and quantification are carried out on the adenosine by calculating a ratio of a fluorescence amount of the microsphere surface after adenosine inflows and a fluorescence amount of the microsphere surface before the adenosine inflows. The adenosine detecting method provided by the invention can be used for detecting and analyzing a target at high sensitivity, so that the specific and accurate quantitative analysis of the adenosine in a serum sample is realized.

Description

technical field [0001] The invention belongs to the technical field of biological detection of a micro-total analysis system, and relates to an adenosine detection method based on a microfluidic chip and nucleic acid aptamer recognition technology. Background technique [0002] Adenosine is an endogenous nucleoside, which has received extensive attention and research because of its important role in the expression of signals in the central nervous system and peripheral nervous system. At the same time, adenosine is the substrate of adenosine deaminase, and its content change will also indirectly reflect the metabolic level of adenosine deaminase in vivo, and the detection of adenosine deaminase activity is an important reference for the diagnosis of many clinical diseases Therefore, the development of new detection methods for adenosine is of great significance for medical clinical diagnosis and body metabolism. Traditional adenosine detection methods mainly include HPLC, u...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/588G01N33/6893
Inventor 张何傅昕胡鑫江
Owner HUNAN INSTITUTE OF ENGINEERING
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