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Electrochemical sensor for detecting ATP and preparation method thereof

A sensor, electrochemical technology, applied in the direction of material electrochemical variables, scientific instruments, instruments, etc., can solve the problems of high cost, low specificity and sensitivity

Active Publication Date: 2019-04-09
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve the problems of relatively low specificity and sensitivity and high cost of the method for detecting ATP in the above prior art, the present invention provides a method based on adjacent induced strand displacement and branching with high specificity and sensitivity, low cost and fast detection speed. Electrochemical sensor for detection of ATP by HCR

Method used

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  • Electrochemical sensor for detecting ATP and preparation method thereof
  • Electrochemical sensor for detecting ATP and preparation method thereof
  • Electrochemical sensor for detecting ATP and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] A preparation method of the electrochemical biosensor of the present invention:

[0072] 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;

[0073] b. Add 10 μL (0 μM, 0.5 μM, 1 μM, 1.5 μM, 2 μM) of CP-SH onto the electrode surface, incubate at room temperature for 2 h, and wash.

[0074] 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:

[0075] a. Add 1X PBS buffer, 1 μM Apt-1, 1 μM Apt-2, 1 μM AP-TP hybrid double strand, 1 μM FP and 2 μL each of the target to be tested, and 8 μL of sterile water into the centrifuge tube , shaken for 30 s, and incubated in a 37°C incubator for 2 h;

[0076] b. Add the solution (10 μL) after the reaction in step a dropwise onto the electrode modified with CP-SH in advance. Then keep the e...

Embodiment 2

[0085] A preparation method of the electrochemical biosensor of the present invention:

[0086] 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;

[0087] b. Add 10 μL of 1 μM CP-SH dropwise to the electrode surface, incubate at room temperature for 2 h, and wash.

[0088] 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:

[0089] a. Add 1X PBS buffer, 1 μM Apt-1, 1 μM Apt-2, 1 μM AP-TP hybrid double strand, 1 μM FP and 2 μL each of the target to be tested, and 8 μL of sterile water into the centrifuge tube , shaken for 30 s, and incubated in a 37°C incubator for 2 h;

[0090] b. Add the solution (10 μL) after the reaction in step a dropwise onto the electrode modified with CP-SH in advance. Then keep the electrode in the incubat...

Embodiment 3

[0099]A preparation method of the electrochemical biosensor of the present invention:

[0100] 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;

[0101] b. Add 10 μL of 1 μM CP-SH dropwise to the electrode surface, incubate at room temperature for 2 h, and wash.

[0102] 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:

[0103] a. Mix 1X PBS buffer, 1 μM Apt-1, 1 μM Apt-2, 1 μM AP-TP hybrid double-strand, 1 μM FP and the target to be tested (0 pM, 1 pM, 10 pM, 100 pM, 1 nM, 10 nM, 100 nM, 1 μM, 5 μM) each 2 μL, sterilized water 8 μL were added to the centrifuge tube, shaken for 30 s, and incubated in a 37 °C incubator for 2 h;

[0104] b. Add the solution (10 μL) after the reaction in step a dropwise onto the electrode modified ...

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Abstract

The invention relates to the technical field of the biosensor, and especially relates to a biosensor for detecting ATP by bringing Apt-1 and Apt-2 closer by ATP and using branch HCR amplification signal. A CP-SH layer, an AP layer, a HP1-HP2-HP3-HP4 layer and a [Ru(NH3)6]3+ layer are orderly modified on an electrode; the preparation method comprises the following steps: pre-treating the electrode,modifying the CP-SH layer to an electrode surface; modifying the AP layer to the electrode surface, and modifying the HP1-HP2-HP3-HP4 layer to the electrode surface, and modifying the[Ru(NH3)6]3+ layer to the electrode surface. The high specificity detection of the target object ATP is realized by utilizing the specificity identification of the aptamer; by utilizing the effect of bring two aptamers closer by the ATP, the cyclic utilization of the target object is realized, and the effect of the first-step signal amplification is achieved. And a signal continuous enhancement effect is realizedby utilizing branch HCR.

Description

technical field [0001] The invention relates to the technical field of electrochemical sensors, in particular to an electrochemical sensor for detecting ATP based on adjacent induced strand displacement and branched HCR, and also relates to a preparation method thereof. Background technique [0002] Adenosine triphosphate (ATP), the most important energy source in cells, plays an important role in intracellular metabolic, synthetic and biochemical processes. Notably, ATP is capable of indicating many human diseases. Typically, abnormal ATP concentrations are closely related to diseases such as Parkinson's, hypoglycemia, tissue hypoxia, ischemia, and cardiovascular diseases. Therefore, it is urgent and important to construct an effective analytical method capable of detecting trace ATP. [0003] At present, commonly used ATP detection methods include mass spectrometry, high performance liquid chromatography, etc. These methods often have problems such as expensive instrumen...

Claims

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Application Information

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IPC IPC(8): G01N27/26G01N27/30G01N27/327
CPCG01N27/26G01N27/30G01N27/3275
Inventor 刘素宋晓蕾黄加栋王玉张雪王敬锋王海旺
Owner UNIV OF JINAN
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