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Method for detecting 8-hydroxy-2 '-deoxyguanosine by using gold nanoparticle-based immunochromatography test paper

An immunochromatographic test paper and gold nanoparticle technology, which is applied in the detection field of 8-hydroxy-2'-deoxyguanosine, can solve the problems of complicated operation, high cost, slow speed, etc., and achieve sensitive analysis and detection, and easy synthesis Effect

Pending Publication Date: 2021-09-28
ZHEJIANG UNIV OF TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] In order to overcome the shortcomings of the existing 8-OHdG detection methods such as slow speed, complicated operation, and high cost, the present invention provides a simple, fast, and sensitive gold nanoparticle-based immunochromatographic test paper for the detection of 8-hydroxyl-2' - The method of deoxyguanosine

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  • Method for detecting 8-hydroxy-2 '-deoxyguanosine by using gold nanoparticle-based immunochromatography test paper
  • Method for detecting 8-hydroxy-2 '-deoxyguanosine by using gold nanoparticle-based immunochromatography test paper
  • Method for detecting 8-hydroxy-2 '-deoxyguanosine by using gold nanoparticle-based immunochromatography test paper

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Embodiment 1

[0033] Embodiment 1: A method for detecting 8-hydroxyl-2'-deoxyguanosine based on gold nanoparticle immunochromatographic test paper, comprising the following steps:

[0034](1) Preparation of gold nanoparticles and surface biofunctionalization treatment: Gold nanoparticles were prepared by reducing perchlorauric acid with sodium citrate, the concentration of the prepared gold nanoparticles was 6nM, and 8-OHdG was modified on the surface of gold nanoparticles The volume ratio of antibody, gold nanoparticles and 8-OHdG antibody (1.2μg / mL) is 5:1, the concentration of the prepared gold-labeled antibody is 96nM, and it is stored at 4°C for later use;

[0035] (2) 8-OHdG complete antigen preparation: prepare 0.96g / L 8-OHdG solution, 4.5g / L bovine serum albumin BSA solution and 28.5g / L carbodiimide hydrochloride EDC solution, 8-OHdG solution and BSA The pH of the solution was adjusted to 5 with dilute hydrochloric acid, and the three solutions were mixed and shaken. The volume rati...

Embodiment 2

[0038] Embodiment 2: A method for detecting 8-hydroxyl-2'-deoxyguanosine based on gold nanoparticle immunochromatographic test paper, comprising the following steps:

[0039] (1) Preparation of gold nanoparticles and surface biofunctionalization treatment: Gold nanoparticles were prepared by reducing perchlorauric acid with sodium citrate, the concentration of the prepared gold nanoparticles was 4nM, and 8-OHdG was modified on the surface of gold nanoparticles The volume ratio of antibody, gold nanoparticles and 8-OHdG antibody (0.5μg / mL) is 5:1, the concentration of the prepared gold-labeled antibody is 40nM, and it is stored at 4°C for later use;

[0040] (2) 8-OHdG complete antigen preparation: prepare 0.96g / L 8-OHdG solution, 4.5g / L bovine serum albumin BSA solution and 28.5g / L carbodiimide hydrochloride EDC solution, 8-OHdG solution and BSA The pH of the solution was adjusted to 5 with dilute hydrochloric acid, and the three solutions were mixed and shaken. The volume rat...

Embodiment 3

[0043] Embodiment 3: A method for detecting 8-hydroxyl-2'-deoxyguanosine based on gold nanoparticles-based immunochromatographic test paper, comprising the following steps:

[0044] (1) Preparation of gold nanoparticles and surface biofunctionalization treatment: Gold nanoparticles were prepared by reducing perchloroauric acid with sodium citrate, the concentration of the prepared gold nanoparticles was 40nM, and 8-OHdG was modified on the surface of gold nanoparticles The volume ratio of antibody, gold nanoparticles and 8-OHdG antibody (1.5μg / mL) is 5:1, the concentration of the prepared gold-labeled antibody is 400nM, and it is stored at 4°C for later use;

[0045] (2) 8-OHdG complete antigen preparation: prepare 0.96g / L 8-OHdG solution, 4.5g / L bovine serum albumin BSA solution and 28.5g / L carbodiimide hydrochloride EDC solution, 8-OHdG solution and BSA The pH of the solution was adjusted to 5 with dilute hydrochloric acid, and the three solutions were mixed and shaken. The ...

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Abstract

The invention relates to a method for detecting 8-hydroxy-2 '-deoxyguanosine by using gold nanoparticle-based immunochromatography test paper. The method comprises the following steps: reducing tetrachloroauric acid trihydrate by sodium citrate to prepare the gold nanoparticles (AuNPs), and coating an 8-OHdG antibody (Ab) on the outer layer of the gold nanoparticles under an alkaline condition to form Ab@AuNPs as a probe; coupling bovine serum albumin (BSA) and 8-OHdG with carbodiimide hydrochloride (EDC) to prepare an artificial antigen, coating a nitrocellulose membrane with the artificial antigen to form a detection line (T line), and coating a nitrocellulose membrane with goat anti-mouse IgG to form a quality control line (C line). The 8-OHdG in the object to be detected and the artificial antigen coated on the detection line compete for the 8-OHdG antibody, so that the 8-OHdG can be detected through the lightening change of the color of the detection line, and the detection limit of naked eyes is 50mg / L. According to the invention, 8-OHdG can be simply, rapidly and sensitively detected.

Description

technical field [0001] The invention relates to a detection technology method of 8-hydroxy-2'-deoxyguanosine, in particular to a method for detecting 8-hydroxy-2'-deoxyguanosine with an immunochromatographic test paper based on gold nanoparticles. Background technique [0002] Reactive oxygen radicals (ROS) can induce DNA oxidative damage in animals and humans, forming a large number of DNA damage products. The presence of phthalates, heavy metals, preservatives, pesticide and veterinary drug residues in food can increase the level of ROS in human cells. In addition, exogenous reactive oxygen species can also cause DNA oxidative damage, such as smoking, ultraviolet radiation and ionizing radiation. 8-OHdG is the most studied DNA oxidative damage product and is recognized as a suitable biomarker of oxidative stress. 8-OHdG has the ability to pair with any base, which can cause gene mutation and oncogene expression during DNA replication. [0003] At present, the commonly u...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/558
CPCG01N33/54346G01N33/558
Inventor 叶伟伟王丽文张宇钱天润
Owner ZHEJIANG UNIV OF TECH
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