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Method for detecting oxidative damage DNA based on fluorescence resonance energy transfer of carbon quantum dot and gold nanoparticle

A technology of fluorescence resonance energy and gold nanoparticles, which is applied in biochemical equipment and methods, and microbial measurement/inspection, etc., can solve problems such as complicated procedures, slow speed, and poor sensitivity, and achieve the effect of sensitive analysis and detection

Active Publication Date: 2017-10-24
ZHEJIANG UNIV OF TECH
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Problems solved by technology

[0005] In order to overcome the shortcomings of the existing methods for detecting damaged DNA, such as slow speed, complicated procedures, and poor sensitivity, the present invention provides a fast, simple, and sensitive fluorescence resonance energy transfer detection oxidation based on carbon quantum dots and gold nanoparticles. Ways to damage DNA

Method used

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  • Method for detecting oxidative damage DNA based on fluorescence resonance energy transfer of carbon quantum dot and gold nanoparticle
  • Method for detecting oxidative damage DNA based on fluorescence resonance energy transfer of carbon quantum dot and gold nanoparticle
  • Method for detecting oxidative damage DNA based on fluorescence resonance energy transfer of carbon quantum dot and gold nanoparticle

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example 1

[0033] Example 1: A method for detecting oxidatively damaged DNA based on fluorescence resonance energy transfer of carbon quantum dots and gold nanoparticles, the process is as follows figure 1 Shown:

[0034] (1) Biofunctionalization treatment of CQDs surface: add 1 μL glutaraldehyde (mass fraction 50%) to 25 μg / mL, 100 μL surface-aminated CQDs solution, let it stand for 30 minutes, then add oxidatively damaged DNA, the concentration range is 1pM~30μM , stored at 4°C for later use;

[0035] (2) AuNPs preparation and surface functionalization treatment: AuNPs were prepared by reducing perchloroauric acid with sodium citrate. Deionized water was heated to boiling on a magnetic stirring heater, 15 μL of perchlorauric acid (14.3%) was added to 50 μL of boiling water, after stirring and heating for 5 minutes, 5 mL of sodium citrate (1%) solution was quickly added, and the solution was carried out under stirring and heating. reaction. The color of the solution gradually changed...

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Abstract

The invention relates to a method for detecting oxidative damage DNA based on fluorescence resonance energy transfer of carbon quantum dot and gold nanoparticle. The method comprises the following steps: (1) bio-functionalization treatment for CQDs surface: adding glutaraldehyde into a surface aminated CQDs solution, standing by for 10-40min, adding oxidative damage DNA, storing at 4 DEG C and standing by; (2) AuNPs preparation and surface bio-functionalization treatment: utilizing a method for reducing high chloroauric acid by using sodium citrate to prepare AuNPs and connecting 8-OHdG antibody with the prepared AuNPs surface through an amino terminal on the antibody, wherein the mass ratio of AuNPs to 8-OHdG antibody is at (1:10)-(1:50); (3) determining the oxidative damage DNA: adding the 8-OHdG antibody modified AuNPs solution into the oxidative damage DNA modified CQDs solution, incubating for 0.5-2 hours and then determining fluorescence intensity. The method is quick, convenient and sensitive.

Description

technical field [0001] The present invention relates to a method for detecting oxidatively damaged DNA, in particular to a method for detecting oxidatively damaged DNA based on fluorescence resonance energy transfer of carbon quantum dots and gold nanoparticles. Background technique [0002] Environmental estrogen has similar properties to biological endocrine hormones and can interfere with the normal physiological functions of the human endocrine system. Common environmental estrogens include bisphenol A, phthalate plasticizers, and synthetic estrogens derived from steroid derivatives similar in structure to estradiol. These environmental pollutants are highly lipophilic or fat-soluble, and can easily enter the human body through the food chain and accumulate in animals and humans. Environmental estrogen is metabolized in the human body to generate active oxygen free radicals such as hydroxyl free radicals and superoxide anions, which attack the 8th carbon atom of the gua...

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6804C12Q2563/107C12Q2565/607C12Q2565/101
Inventor 叶伟伟陈畑毛燚杰孙培龙
Owner ZHEJIANG UNIV OF TECH
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