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Preparation method and application of a fluorescent biosensor

A biosensor and fluorescence technology, which is applied in the field of fluorescent biosensors, detection of HBV gene, and preparation of fluorescent biosensors, can solve the problems of time-consuming and lengthy detection process, complexity, etc., to avoid chemical labeling and modification, and to achieve low detection limit , the effect of simple operation

Active Publication Date: 2017-12-15
ANHUI NORMAL UNIV
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Problems solved by technology

[0004] At present, the detection of HBV genes in actual samples mostly involves complex and time-consuming detection processes, or involves the labeling of external fluorescent signal molecules, so the development of highly selective, high-sensitivity, simple and label-free fluorescent biosensor detection HBV Genes Are Crucial

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  • Preparation method and application of a fluorescent biosensor
  • Preparation method and application of a fluorescent biosensor
  • Preparation method and application of a fluorescent biosensor

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Embodiment 1

[0045] Verify that G-quadruplexes can quench Cu NPs:

[0046]a. Add 1 μL of polyT30 solution to 80 μL of 10 mM MOPS (3-(N-morpholino)propanesulfonic acid) buffer solution at pH=7.6, then add 5 μL of CuSO4 solution (10 mM) and 120 μL of sodium ascorbate solution (100mM), to obtain mixed solution 1, to obtain fluorescent Cu NPs, and to detect the fluorescence intensity;

[0047] The preparation method of the polyT30 solution is as follows: polyT30 is dissolved in a buffer solution of 10 mM MOPS (3-(N-morpholino)propanesulfonic acid) pH=7.6, so that the final concentration of the polyT30 sequence is 500 nM.

[0048] b. Take 1 μL of G4 solution (500 nM), 0.5 μL of hemin solution (1.0 μM) and 5 μL of KCl solution (50 mM) into 200 μL of MOPS (10 mM MOPS (3-(N-morpholino)propanesulfonic acid) PH =7.6 buffer solution), after cultivating in a dry box at 37°C for 2 hours, a mixed solution 2 was obtained; (G4 gene sequence and hemin and K + Mix together to form a G-quadruplex structure...

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Abstract

The invention discloses a method for preparing a fluorescent biosensor and its application. Copper nanoparticles carrying polythymine single-stranded DNA as a template are used for luminescence, and light-induced electron transfer is generated between G-quadruples and copper nanoparticles. The luminescence of copper nanoparticles is quenched, and a fluorescent sensor is constructed based on copper nanoparticles carrying thymine DNA as a template and a light-induced electron transfer system. The combination of the target (HBV gene) and the probe forms a compact system The principle of extending the distance between G-quadruples and copper nanoparticles realizes the detection of HBV gene sensitivity and specificity.

Description

technical field [0001] The invention belongs to the technical field of fluorescent sensor preparation, and specifically relates to a method for preparing a fluorescent biosensor and its application. The fluorescent biosensor constructed by the electron transfer system is used in the detection of HBV gene. Background technique [0002] Hepatitis B virus (HBV), a DNA virus that can be transmitted through blood, semen and other bodily fluids, is one of the most common and highly contagious infectious diseases worldwide and can cause chronic hepatitis, cirrhosis and primary liver cancer. Hepatitis B virus (HBV) belongs to the hepadnaviridae family, and its genome is about 3.2kb in length, which is partially double-stranded circular DNA. [0003] About two billion people in the world have been infected with this virus, and about 350 million people are in the infection stage. All countries are facing challenges and threats caused by HBV virus infection. Early diagnosis of the fa...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64
Inventor 王广凤万靖
Owner ANHUI NORMAL UNIV
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