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Method for preparing fluorescence biosensor and application of fluorescence biosensor

A biosensor, fluorescence technology, applied in the field of fluorescent biosensor, application of detecting HBV gene, and preparation of fluorescent biosensor, can solve the problems of complex, time-consuming and lengthy detection process, achieve low detection limit, avoid chemical labeling and modification , the effect of simple operation

Active Publication Date: 2015-12-30
ANHUI NORMAL UNIV
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  • Abstract
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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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  • Method for preparing fluorescence biosensor and application of fluorescence biosensor
  • Method for preparing fluorescence biosensor and application of fluorescence biosensor
  • Method for preparing fluorescence biosensor and application of fluorescence biosensor

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Embodiment 1

[0045] Verify that G-quadruplexes can quench CuNPs:

[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 CuNPs, and to detect the fluorescence intensity;

[0047] The preparation method of the polyT30 solution is as follows: dissolving polyT30 into 10 mM MOPS (3-(N-morpholino)propanesulfonic acid) buffer solution at pH=7.6, so that the final concentration of 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 culturing 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, and ...

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Abstract

The invention discloses a method for preparing a fluorescence biosensor and an application of the fluorescence biosensor. Under the light-emitting effect of copper nano particles with polythymine-carried single-stranded DNAs used as the template, the phenomenon that light induction electron transfer is generated between G-four-linked bodies and the copper nano particles, and thus emitted light of the copper nano particles is quenched is utilized, so that the copper nano particles based on the thymine-carried single-stranded DNAs used as the template and a light induction electron transfer system are prepared for establishing the fluorescence sensor. The principle that targets (HBV genes) and probes are combined to form the system of a compact structure, so that the distance between the G-four-linked bodies and the copper nano particles is zoomed out is adopted, and the sensitivity and specificity of the HBV genes are detected.

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 Applications(China)
IPC IPC(8): G01N21/64
Inventor 王广凤万靖
Owner ANHUI NORMAL UNIV
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