Aptamer-based biosensor for detecting penbritin and preparation method of aptamer-based biosensor

A biosensor, ampicillin 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, improved sensitivity and accurate quantitative detection

Inactive Publication Date: 2015-03-25
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 ampicillin in the above prior art, the present invention provides a nucleic acid aptamer-based detection method with high specificity and sensitivity, low cost and fast detection speed. Ampicillin biosensor

Method used

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  • Aptamer-based biosensor for detecting penbritin and preparation method of aptamer-based biosensor
  • Aptamer-based biosensor for detecting penbritin and preparation method of aptamer-based biosensor
  • Aptamer-based biosensor for detecting penbritin and preparation method of aptamer-based biosensor

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] The main steps of the electrode modification process are as follows:

[0056] a. The gold electrode is firstly polished in 0.3 and 0.05??m alumina slurry until it becomes a mirror surface, and then rinsed repeatedly with PBS and secondary water;

[0057] b. Drop 10 μL of capture probe (10 μM) onto the electrode surface and incubate at room temperature (25°C) for 2 hours. Fix the sulfhydryl chains to the electrode surface through Au-S bonds;

[0058] 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:

[0059] a. Add sterilized water, 10X buffer (buffer), 1 μL hairpin DNA strands (concentrations are 20 μM, 10 μM, 5 μM, 2 μM, 1 μM) and the target substance to a pre-prepared sterilized centrifuge tube middle. Shake for 30s, then put the mixed mixture into a 37°C incubator and incubate for 2h;

[0060] b. Continue to add MB probe (10 μM), po...

Embodiment 2

[0070] A preparation method of the electrochemical biosensor of the present invention:

[0071] The main steps of the electrode modification process are as follows:

[0072] a. The gold electrode is firstly polished in 0.3 and 0.05??m alumina slurry until it becomes a mirror surface, and then rinsed repeatedly with PBS and secondary water;

[0073] b. Drop 10 μL of capture probe (10 μM) onto the electrode surface and incubate at room temperature for 2 hours. Fix the sulfhydryl chains to the electrode surface through Au-S bonds;

[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 sterilized water, 10X buffer, hairpin-structured DNA strand (10 μM) and the target substance into a pre-prepared sterilized centrifuge tube. Shake for 30s, then put the mixed mixture into a 37°C incubator and incubate for 2h;

[0076] b. Continu...

Embodiment 3

[0082] A preparation method of the electrochemical biosensor of the present invention:

[0083] a. The gold electrode is firstly polished in 0.3 and 0.05??m alumina slurry until it becomes a mirror surface, and then rinsed repeatedly with PBS and secondary water;

[0084] b. Add 10 μL of different concentrations of capture probe (20 μM, 10 μM, 5 μM, 2 μM, 1 μM) onto the electrode surface and incubate at room temperature for 2 hours. Fix the sulfhydryl chains to the electrode surface through Au-S bonds;

[0085] 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:

[0086] a. Add sterilized water, 10X buffer, hairpin-structured DNA strand (10 μM) and the target substance into a pre-prepared sterilized centrifuge tube. Shake for 30s, then put the mixed mixture into a 37°C incubator and incubate for 2h;

[0087] b. Continue to add MB probe (10 μM), ...

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Abstract

The invention relates to the technical field of a biosensor, and particularly relates to an aptamer-based biosensor for detecting penbritin, which is used for solving the problems of low specificity and sensitivity in the traditional method for detecting the penbritin in the prior art. The aptamer-based biosensor for detecting the penbritin is characterized in that a capture probe layer, HAP and an MB probe layer are sequentially decorated on an electrode. The preparation method comprises the following steps: preprocessing the electrode; decorating the capture probe layer onto the surface of the electrode; decorating the HAP and the MB probe layer onto the surface of the electrode. By utilizing the specificity recognition of aptamer, the aptamer of penbritin is used as a recognition substance, so that the high-specificity detection on the target penbritin is realized; by utilizing the polymerization effect of polymerase, the cycling utilization of a target substance is realized, and an effect on amplifying the signal is realized.

Description

technical field [0001] The invention relates to the technical field of biosensors, in particular to a biosensor for detecting ampicillin based on nucleic acid aptamers, and also relates to a preparation method thereof. Background technique [0002] Ampicillin is a broad-spectrum semi-synthetic penicillin with a similar antibacterial spectrum to penicillin. It has been widely used in medicine and agriculture to treat bacterial infections. He can effectively treat many bacteria, such as Haemophilus influenzae, Escherichia coli, Salmonella, Shigella, etc. However, the drug residues in food and agricultural products caused by the abuse of ampicillin in animal husbandry and the diseases caused by these residues, such as: allergic reactions, dyspnea and seizures, etc. Therefore, determining the residues of ampicillin in food and agricultural products has become very important. At present, institutions including the US Food and Drug Administration (FDA), the Joint Committee...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/573
CPCG01N33/573
Inventor 黄加栋王虹智王玉刘素崔敏李杰徐伟郭玉娜许颖崔杰邱婷婷
Owner UNIV OF JINAN
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