Preparation method of electrochemical DNA biosensor for detection of rs1801177

A biosensor and electrochemical technology, applied in the field of electrochemical detection, can solve the problem that DNA molecules do not have electrochemical activity, etc., and achieve the effects of improving sensitivity and biocompatibility, fewer detection steps, and faster electron transfer.

Inactive Publication Date: 2019-03-29
CHONGQING MEDICAL UNIVERSITY
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  • Abstract
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  • Claims
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Problems solved by technology

Although the nanocomposite material formed by nitrogen-doped graphene and palladium-platinum alloy does help to amplify the electrical signal and enhance the sensitivity, the DNA molecule itself is not electrochemically active.

Method used

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  • Preparation method of electrochemical DNA biosensor for detection of rs1801177
  • Preparation method of electrochemical DNA biosensor for detection of rs1801177
  • Preparation method of electrochemical DNA biosensor for detection of rs1801177

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Embodiment 1

[0042] Step 1.C 60 / PAMAM nanocomposite material is prepared by the method of phase transfer, and its specific operation steps are as follows: Weigh 1.08mg C 60 powder, add 1mL of toluene and sonicate for 30min to completely disperse and form a uniform purple solution. Under the condition of magnetic stirring, transfer it into 8mL small beaker, add 25uL PAMAM, 1mL ultrapure water, 4m ethanol. After reacting for 36 hours, centrifuge and wash several times to remove excess substances, and then disperse it in 2mL ultrapure water;

[0043] Step 2. Under constant stirring, 400 μL of chloroauric acid (1%) was added to the above synthesized 2mL C 60 / PAMAM nanocomposite solution, followed by 4mL sodium borohydride (2mg mL -1 ) solution was added dropwise to the mixed solution, and after stirring at room temperature for half an hour, it was centrifuged and washed, and the obtained C 60 / PAMAM / Au nanocomposites were redispersed in 2mL ultrapure water and stored at 4°C for use;

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Abstract

The invention relates to a preparation method of an electrochemical DNA biosensor for detection of LPL gene SNP (rs1801177), and belongs to the technical field of electrochemical detection. The preparation method is characterized by comprising the steps: firstly, the surface of a nitrogen-doped graphene (N-G) sheet is loaded with a large number of palladium-platinum bimetal (PdPt) nanoparticles, then polyaniline is formed on the surface of the nitrogen-doped graphene (N-G) sheet by in-situ polymerization to obtain a nano composite material, next, a single-stranded DNA signal probe is mixed with the composite material, and a redox probe is prepared; secondly, a capture probe is immobilized on the surface of a sensor by a C60 / PAMAM / Au nanocomposite material, and the electrochemical DNA biosensor for quantitative detection of rs1801177 is prepared. The biosensor is successfully applied to detection of single-base mutation of a lipoprotein lipase gene. The biosensor has the advantages of high sensitivity, strong specificity, and convenient and fast detection, provides a new method for detection of rs1801177, and provides a theoretical basis and an experimental basis for clinical diagnosis and prevention of cardiovascular diseases.

Description

Technical field: [0001] The present invention relates to a preparation method and application of an electrochemical immunosensor for LPL gene SNP (rs1801177) detection, especially a sandwich-type electrochemical DNA biosensor based on nitrogen-doped graphene nanocomposite materials for the detection of rs1801177, It belongs to the field of electrochemical detection. Background technique: [0002] Cardiovascular disease (CVD) has a high incidence worldwide and seriously endangers human health. Atherosclerosis (AS) is the pathological basis of cardiovascular diseases and one of the important causes of death of patients. Early identification and intervention of atherosclerosis is an important means to reduce the morbidity and mortality of cardiovascular diseases. Studies have shown that the lipoprotein lipase (Lipoprotein lipase, LPL) gene consists of 10 exons and 9 introns, the total length is about 36kb, and the coding region of the peptide is about 23kb. There is single n...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/44C12Q1/00G01N27/48G01N27/327
CPCG01N27/3278G01N27/48C12Q1/005C12Q1/44
Inventor 于超周媛吴静
Owner CHONGQING MEDICAL UNIVERSITY
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