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A kind of non-labeled nucleic acid aptamer sensor and its detection method for bisphenol A

A nucleic acid aptamer and detection method technology, applied in the field of chemical sensing and electroanalytical chemical detection, can solve the problems of affecting the binding affinity of aptamers and target molecules, complex labeling process, and high cost

Inactive Publication Date: 2018-04-03
EAST CHINA JIAOTONG UNIVERSITY
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Electrochemical nucleic acid aptamer sensors based on nucleic acid aptamers can be divided into labeled and non-labeled types according to whether markers are used to generate detection signals; the labeling process of labeled nucleic acid aptamer sensors is complicated and expensive, and in To some extent, it will affect the binding affinity of the aptamer and the target molecule

Method used

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  • A kind of non-labeled nucleic acid aptamer sensor and its detection method for bisphenol A
  • A kind of non-labeled nucleic acid aptamer sensor and its detection method for bisphenol A
  • A kind of non-labeled nucleic acid aptamer sensor and its detection method for bisphenol A

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Preparation of nucleic acid aptamer sensors based on porous gold nanoparticles and methylene blue electrochemical probes, such as figure 1 shown.

[0026] (1) Prepare 0.04 mol / L HAuCl 4 and 0.002 mol / L of Pb(CH 3 COO) 2 solution, the clean glassy carbon electrode was placed in the above solution, and was electro-deposited at -0.2 V for 120 s at a constant potential to prepare a porous nano-gold modified glassy carbon electrode.

[0027] (2) The porous nano-gold modified electrode was placed in 0.3 μmol / L bisphenol A aptamer for 24 h, 0.5 mmol / L 6-mercaptohexanol for 80 min, and 0.3 μmol / L complementary DNA for 80 min. min, the double-stranded DNA / porous gold nano-modified glassy carbon electrode was prepared.

[0028] (3) Soak the glassy carbon electrode modified with double-stranded DNA / porous gold nanoparticles in 0.1 mmol / L methylene blue solution for 25 min to obtain a glassy carbon electrode modified with methylene blue / double-stranded DNA / porous gold nanoparti...

Embodiment 2

[0030] Preparation of nucleic acid aptamer sensors based on porous gold nanoparticles and methylene blue electrochemical probes, such as figure 1 shown.

[0031] (1) Prepare 0.08 mol / L HAuCl 4 and 0.004 mol / L of Pb(CH 3 COO) 2 solution, a clean glassy carbon electrode was placed in the above solution, and was electro-deposited at a constant potential of -0.5 V for 70 s to prepare a porous nano-gold modified glassy carbon electrode.

[0032] (2) The porous nano-gold modified electrode was placed in 1.0 μmol / L bisphenol A aptamer for 12 h, 1.0 mmol / L 6-mercaptohexanol for 60 min, and 1.0 μmol / L complementary DNA for 60 min. min, the double-stranded DNA / porous gold nano-modified glassy carbon electrode was prepared.

[0033] (3) Soak the glassy carbon electrode modified with double-stranded DNA / porous gold nanoparticles in 0.2 mmol / L methylene blue solution for 10 min to obtain a glassy carbon electrode modified with methylene blue / double-stranded DNA / porous gold nanoparticle...

Embodiment 3

[0035] Preparation of nucleic acid aptamer sensors based on porous gold nanoparticles and methylene blue electrochemical probes, such as figure 1 shown.

[0036] (1) Prepare 0.12 mol / L HAuCl 4 and 0.006 mol / L of Pb(CH 3 COO) 2 solution, the clean glassy carbon electrode was placed in the above solution, and was electro-deposited at -0.7 V for 20 s at a constant potential to prepare a porous nano-gold modified glassy carbon electrode.

[0037] (2) The porous nano-gold modified electrode was placed in 1.5 μmol / L bisphenol A aptamer for 6 hours, 1.5 mmol / L 6-mercaptohexanol for 20 minutes, and 1.5 μmol / L complementary DNA for 20 minutes. min, the double-stranded DNA / porous gold nano-modified glassy carbon electrode was prepared.

[0038](3) Soak the glassy carbon electrode modified with double-stranded DNA / porous gold nanoparticles in 0.4 mmol / L methylene blue solution for 5 min to obtain the glassy carbon electrode modified with methylene blue / double-stranded DNA / porous gold...

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Abstract

A non-labeled nucleic acid aptamer sensor and a detection method for bisphenol A. The method of the present invention uses porous nano-gold with a large specific surface area as the base material of the immobilized nucleic acid aptamer, and then hybridizes the loaded nucleic acid aptamer Complementary DNA strands formed double-stranded DNA, and finally the electrochemical probe methylene blue was embedded to construct a non-labeled nucleic acid aptamer electrochemical sensor for the determination of bisphenol A. The sensitization effect of the porous nano-gold and the indication effect of the methylene blue electrochemical probe can detect the bisphenol A sensitively, accurately and specifically. The invention only needs to change the nucleic acid aptamer and the complementary DNA chain, and can realize the detection of other substrates. The invention is suitable for measuring bisphenol A by an electrochemical nucleic acid aptamer sensor method.

Description

technical field [0001] The invention relates to a non-labeled nucleic acid aptamer sensor and a detection method for bisphenol A, belonging to the technical fields of chemical sensing and electroanalysis chemical detection. Background technique [0002] Bisphenol A is one of the environmental estrogen compounds, which can enter the body to imitate, hinder, interfere with or change the body's own hormone action, leading to endocrine system disorders, hindering the transmission of the nervous system, reducing the body's immune function, and even causing organ deformities and Cancer is a serious threat to human health and survival. Therefore, the rapid and accurate detection of bisphenol A has important practical significance. [0003] Nucleic acid aptamer is a single-stranded or double-stranded oligonucleotide composed of 20-60 bases screened in vitro by the exponential enrichment ligand system evolution technology, with high affinity, easy modification and function It has t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/327
CPCG01N27/3277
Inventor 杨绍明郑玥李玲玲张小荣丁绍卿陈爱喜
Owner EAST CHINA JIAOTONG UNIVERSITY
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