Preparation method and application of electrochemical biosensor based on DNA walker

A biosensor and electrochemical technology, applied in the field of biosensors, can solve the problems of insufficient sensitivity and accuracy, and be easily affected by environmental factors, so as to achieve the effect of improving sensitivity and simple construction

Pending Publication Date: 2021-01-29
JIANGSU UNIV
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Problems solved by technology

Various electrochemical sensors for the sensitive detection of AF have been constructed, such as a multifunctional DNA nanotube-based electrochemical sensor for AFB1 analysis, and dual-mode sensors combined by conventional electrochemical methods; however, due to the single-signal output Susceptible to environmental factors and lacks sensitivity and precision

Method used

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  • Preparation method and application of electrochemical biosensor based on DNA walker
  • Preparation method and application of electrochemical biosensor based on DNA walker
  • Preparation method and application of electrochemical biosensor based on DNA walker

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Embodiment 1

[0054] (1) Mix the DNA walker solution with a concentration of 0.1 μM, the hairpin DNA1 solution at 5 μM, and the hairpin DNA2 solution at 5 μM for a reaction time of 5 minutes in a water bath at 95 ° C. After the reaction in the water bath, cool to room temperature to obtain DNAwalker-AFB1 aptamer solution;

[0055] (2) Then the DNA walker-AFB1 aptamer solution was mixed with the hairpin DNA1 solution and the hairpin DNA2 solution to obtain a mixed solution A; then a total of 6 μL of the mixed solution A was modified on the surface of the gold electrode and incubated at 4 °C for 12 hours; A total of 8 μL of 1 mM mercaptohexanol (MCH) was dropped onto the surface of the Au electrode to block the Au active sites not bound to the DNA strand in step 1, and block at room temperature for 1 hour to obtain an electrochemical biosensor based on DNA walker;

[0056] (3) Take 4 electrochemical biosensors that have been constructed in the above steps, and modify the surface with a concen...

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Abstract

The invention belongs to the technical field of biosensors, and relates to a preparation method and application of an electrochemical biosensor based on DNA walker. The method comprises the followingsteps: firstly, mixing a DNA walkerAFB1 aptamer with hairpin DNA1 and hairpin DNA2, dropwise adding a mixture to the surface of a gold electrode, performing reacting, dropwise adding MCH, and performing incubating at normal temperature to obtain an electrochemical biosensor; then, modifying the surface of the biosensor with AFB1, anchoring a mixed solution of a probe 1, short-chain DNA1 and short-chain DNA 2, soaking the biosensor into a methylene blue solution, and constructing a standard curve by measuring the current value of a sensor and the concentration of the AFB1; and finally, measuring the current value of an unknown sample, and substituting the current value into the constructed standard curve so as to achieve the detection of the AFB1 in the unknown sample. According to the invention, dual signal amplification can be carried out by using an extremely small amount of target AFB1 to obtain a larger methylene blue electrochemical signal, and the method has good detection selectivity, high speed and high sensitivity.

Description

technical field [0001] The invention belongs to the technical field of biosensors, and in particular relates to a preparation method and application of a DNA walker-based electrochemical biosensor, which can be used for detecting mycotoxin aflatoxin B1 (AFB1). Background technique [0002] Aflatoxins (AFs) are furan-ring toxins that are widely present in agricultural production, processing and transportation. Mycotoxins such as AFs are classified as class I human carcinogens due to their high hepatotoxicity and carcinogenicity to humans. Many countries and world organizations have put forward strict requirements for AF, such as the maximum residue limit of AF in food is 15ng mL -1 . Among them, aflatoxin B1 (AFB1) has been proved to be extremely dangerous. And developed many recognized advanced analytical techniques to detect AFB1, such as high-performance liquid chromatography-fluorescence detection (HPLC-FL) and enzyme-linked immunosorbent assay (ELISA). However, these...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6825G01N33/53G01N27/327G01N27/416
CPCC12Q1/6825G01N27/327G01N27/4163G01N33/5308C12Q2525/205C12Q2525/301C12Q2565/607
Inventor 由天艳贾帆刘东孟淑云
Owner JIANGSU UNIV
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