Electrochemical sensor for detecting mercury ions based on aptamer and preparation method thereof

A nucleic acid aptamer and electrochemical technology, applied in the field of biosensors, can solve the problems of high cost, low specificity and sensitivity, and achieve the effects of low detection limit, simple preparation method, and accurate quantitative detection.

Inactive Publication Date: 2017-06-13
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the problems of relatively low specificity and sensitivity and high cost of the method for detecting mercury ions in the above prior art, the present invention provides a nucleic acid aptamer-based conformational method with high specificity and sensitivity, low cost and fast detection speed. Change-detection electrochemical sensor for mercury ions

Method used

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  • Electrochemical sensor for detecting mercury ions based on aptamer and preparation method thereof
  • Electrochemical sensor for detecting mercury ions based on aptamer and preparation method thereof
  • Electrochemical sensor for detecting mercury ions based on aptamer and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] The main steps of the electrode modification process are as follows:

[0053] a. The gold electrode is first polished in 0.3 and 0.05 µm alumina slurry until it becomes a mirror surface, and then rinsed repeatedly with PBS and secondary water;

[0054] b. Add 10 μL of HAP (10 μM) dropwise to the electrode surface and incubate at room temperature (37°C) for 2 hours. Fix the sulfhydryl chains to the electrode surface through Au-S bonds;

[0055] So far, the modification process of the electrode has come to an end. The following describes the reaction in the homogeneous solution and the main steps in the homogeneous reaction:

[0056] a. Sterilized water, 1X buffer (buffer), 1 μL Helper1 (concentrations are 1 μM, 2 μM, 5 μM, 10 μM, 20 μM), 1 μL Helper2, restriction endonuclease Nt. BbvCI (1 μL) and target Add to the pre-prepared sterilized centrifuge tube. Shake for 30s, then put the mixed mixture into a 37°C incubator and incubate for 2h;

[0057] b. Drop the solution...

Embodiment 2

[0066] A preparation method of the electrochemical biosensor of the present invention:

[0067] The main steps of the electrode modification process are as follows:

[0068] a. The gold electrode is first polished in 0.3 and 0.05 µm alumina slurry until it becomes a mirror surface, and then rinsed repeatedly with PBS and secondary water;

[0069] b. Add 10 μL of HAP (10 μM) dropwise to the electrode surface and incubate at room temperature for 2 hours. Fix the sulfhydryl chains to the electrode surface through Au-S bonds;

[0070] So far, the modification process of the electrode has come to an end. The following describes the reaction in the homogeneous solution and the main steps in the homogeneous reaction:

[0071] a. Sterilized water, 1X buffer (buffer), 1 μL Helper1, 1 μL different concentrations of Helper2 (concentrations are 1 μM, 2 μM, 5 μM, 10 μM, 20 μM), restriction endonuclease Nt. BbvCI (1 μL) Add the target substance into the pre-prepared sterilized centrifuge...

Embodiment 3

[0077] A preparation method of the electrochemical biosensor of the present invention:

[0078] a. The gold electrode is first polished in 0.3 and 0.05 µm alumina slurry until it becomes a mirror surface, and then rinsed repeatedly with PBS and secondary water;

[0079] b. Add 10 μL of different concentrations of HAP (1 μM, 2 μM, 5 μM, 10 μM, 20 μM) onto the electrode surface and incubate at room temperature for 2 h. Fix the sulfhydryl chains to the electrode surface through Au-S bonds;

[0080] So far, the modification process of the electrode has come to an end. The following describes the reaction in the homogeneous solution and the main steps in the homogeneous reaction:

[0081] a. Add sterilized water, 1X buffer, Helper1 (10μM), Helper2 (10μM), restriction endonuclease Nt. tube. Shake for 30s, then put the mixed mixture into a 37°C incubator and incubate for 2h;

[0082] b. Drop the solution after the reaction in step a onto the electrode modified with HAP in advance...

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Abstract

The invention provides an electrochemical sensor for detecting mercury ions based on aptamer conformation change. The electrochemical sensor comprises an electrode, wherein the electrode is successively modified with an HAP layer and a Helper layer; the invention also provides a biological preparation method based on specific identification of the aptamer. When the electrochemical sensor provided by the invention is applied to detection, the electrochemical sensor has the characteristics of high specificity, high sensitivity, low cost and rapid detection speed.

Description

technical field [0001] The invention relates to the technical field of biosensors, in particular to an electrochemical sensor for mercury ions based on conformational changes of nucleic acid aptamers, and also relates to a preparation method thereof. Background technique [0002] Mercury and its compounds have many kinds of harm to human health, and if they exist in natural water bodies, they will pose a threat to a wide range of people. It can accumulate in organisms and transfer to the human body through the food chain. The trace mercury accumulated in the human body cannot be excreted through its own metabolism, which will directly lead to heart, liver and thyroid diseases, nervous system disorders, chronic mercury poisoning, and even the formation of malignant tumors. [0003] Dissolved Hg² + It often has high chemical activity and is the main form of mercury pollutants discharged into natural water bodies. Its compounds have high water solubility and are also the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327
CPCG01N27/3275
Inventor 李慧韩聪刘素王玉黄加栋
Owner UNIV OF JINAN
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