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Fluorescence enhancement DNA detection method based on metal nano-particle coupling

A metal nanoparticle and gold nanoparticle technology, which is applied in the field of fluorescence enhanced detection of DNA, can solve the problems of high background, low sensitivity and long time consumption.

Active Publication Date: 2018-07-06
QINGDAO UNIV
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0003] Aiming at the limitations of the existing DNA fragment detection technology, such as high background, low sensitivity, high detection limit, long time consumption, poor stability, etc., the problem to be solved in the present invention is to provide a method for fluorescence enhancement to achieve high detection of DNA. Selective, highly sensitive detection

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  • Fluorescence enhancement DNA detection method based on metal nano-particle coupling
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  • Fluorescence enhancement DNA detection method based on metal nano-particle coupling

Examples

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Embodiment 1

[0026] Step 1: Preparation of 27nm gold nanoparticles: Take 100mL milli-Q water in a three-necked bottle, place it in an oil bath, heat it with magnetic stirring and reflux, add 2mL 1wt% HAuCl to it in turn after boiling 4 And 3mL sodium citrate solution (30mM), the solution color quickly changed from light yellow to light gray, blue, and finally turned into wine red. Continue heating for 20 min, stir and cool to room temperature to obtain 27 nm gold nanoparticles.

[0027] Step 2: Preparation of silver-coated gold nanoparticles by seed growth method: 1) Take 100mL milli-Q water in a three-necked bottle, place it in an oil bath, heat it with magnetic stirring and reflux, add 2mL 1wt% HAuCl to it in turn after boiling 4 and 3.5 mL of sodium citrate solution (30 mM), continued heating for 20 min, stirred and cooled to room temperature to obtain 17 nm gold nanoparticles. 2) Add 3 mL of 17 nm gold nanoparticle solution into 27 mL of water containing 4 mM sodium citrate and 1 mM a...

Embodiment 2

[0034] Preparation of 27nm gold nanoparticles: same as in Example 1.

[0035] Seed growth method to prepare silver-coated gold nanoparticles: 1) The preparation method of gold nanoparticles with a particle size of 17nm is the same as that in Example 1. 2) Get 3mL of 17nm gold nanoparticles solution and add it to 27mL of water containing 4mM sodium citrate and 1mM ascorbic acid , and adjust the pH of the solution to neutral with 0.01M NaOH. Add 3 mL of AgNO at a concentration of 18 mM dropwise within 30 min3 aqueous solution, and then placed in an oil bath and heated to reflux for 1 hour to obtain 65nm silver-coated gold nanoparticles.

[0036] The method for preparing metal nanoparticles modified with DNA chains is the same as above.

[0037] The DNA-modified silver nanoparticles and gold nanoparticles were mixed at a ratio of 2:1, and the fluorescence intensity was measured. After adding different concentrations of target DNA, the fluorescence signal was measured again, and ...

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Abstract

The invention discloses a fluorescence enhancement DNA detection method based on metal nano-particle coupling. The method comprises the following steps: adding a target DNA to a solution containing DNA-modified gold nano-particles and silver-coated gold nano-particles, the gold nano-particles and the silver-coated gold nano-particles are aggregated due to the hydrogen bond interaction between complementary DNA chains; the aggregation enables an electromagnetic field between the nano-particles to be enhanced and the surface plasma absorption to move, thereby enhancing a specific fluorescent molecular fluorescence signal which is used for detecting the DNA. The method is simple in treating process, low in background and high in sensitivity, and is capable of fundamentally eliminating the influence to the target object detection caused by the fluorescence signal in high background.

Description

technical field [0001] The invention relates to a method for detecting DNA based on fluorescence enhancement coupled with metal nanoparticles. Background technique [0002] DNA is the basic unit of genetic information, and changes in the DNA sequence often lead to disease. Therefore, the detection of specific DNA sequences is of great significance for the early diagnosis and prevention of diseases. At present, common DNA detection methods include colorimetric methods based on the aggregation of gold and silver nanoparticles, traditional fluorescence methods, and electrochemical methods. However, these methods have disadvantages such as high background, low detection sensitivity, high detection limit, and poor repeatability. Contents of the invention [0003] Aiming at the limitations of the existing DNA fragment detection technology, such as high background, low sensitivity, high detection limit, long time consumption, poor stability, etc., the problem to be solved in th...

Claims

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Application Information

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IPC IPC(8): G01N21/64
CPCG01N21/6428G01N2021/6432G01N2021/6439
Inventor 朱志军唐建国黄林军刘继宪焦吉庆王光辉
Owner QINGDAO UNIV
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