Label-free aptamer sensor and method for detecting 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: 2016-07-20
EAST CHINA JIAOTONG UNIVERSITY
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  • Summary
  • Abstract
  • 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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  • Label-free aptamer sensor and method for detecting bisphenol A
  • Label-free aptamer sensor and method for detecting bisphenol A
  • Label-free aptamer sensor and method for detecting bisphenol A

Examples

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 0.04mol / L HAuCl 4 and 0.002mol / L of Pb(CH 3 COO) 2 solution, a clean glassy carbon electrode was placed in the above solution, and electrodeposited at a constant potential of -0.2V for 120s 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 hours, 0.5 mmol / L 6-mercaptohexanol for 80 minutes, and 0.3 μmol / L complementary DNA for 80 minutes to prepare A double-stranded DNA / porous gold nano-modified glassy carbon electrode was obtained.

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

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 0.08mol / L HAuCl 4 and 0.004mol / L of Pb(CH 3 COO) 2 solution, a clean glassy carbon electrode was placed in the above solution, and electro-deposited at a constant potential of -0.5V for 70s 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 hours, 1.0 mmol / L 6-mercaptohexanol for 60 minutes, and 1.0 μmol / L complementary DNA for 60 minutes to prepare A double-stranded DNA / porous gold nano-modified glassy carbon electrode was obtained.

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

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

[0037] (2) Place the porous nano-gold modified electrode 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 to prepare A double-stranded DNA / porous gold nano-modified glassy carbon electrode was obtained.

[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 minutes to obtain a glassy carbon electrode modified with methylene blue / double-stranded DNA / porous gold nanoparti...

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Abstract

The invention discloses a label-free aptamer sensor and a method for detecting bisphenol A. According to the method, porous nano-gold with a large specific surface area serves as a base material of immobilized aptamer; then, porous nano-gold is hybridized with a complementary DNA strand loaded with aptamer to form double-strand DNA; finally, double-strand DNA is embedded into a methylene blue electrochemical probe, and thus the label-free aptamer electrochemical sensor for measuring bisphenol A is constructed. With the sensibilization of porous nano-gold and the indicating function of the methylene blue electrochemical probe, bisphenol A can be flexibly, accurately and specifically detected. The label-free aptamer sensor can be used for detecting other substrates just by changing the aptamer and the complementary DNA strand. The label-free aptamer sensor is suitable for measuring bisphenol A through an electrochemical 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 Applications(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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