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An electrochemical aptamer sensor for ampicillin detection

An aptamer sensor and ampicillin technology, applied in the field of biosensors, can solve the problems of low specificity, sensitivity, and high cost, and achieve the effects of improved sensitivity, simple electrodes, and mild reaction conditions

Active Publication Date: 2022-07-29
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In order to solve the problem of low specificity and sensitivity and high cost of the prior art method for detecting ampicillin, the present invention provides an ampicillin electrochemical sensor with high specificity and sensitivity, low cost and fast detection speed. The chain cycle and the DNA walker on the electrode are used to increase the signal, which greatly improves the detection sensitivity and obtains a lower detection limit, which can be used to detect trace amounts of ampicillin in food

Method used

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  • An electrochemical aptamer sensor for ampicillin detection
  • An electrochemical aptamer sensor for ampicillin detection
  • An electrochemical aptamer sensor for ampicillin detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 Preparation of Apt-T Hybrid Chain

[0039] 2 μL of 100 μM aptamer Apt, 2 μL of 100 μM T chain, 2 μL 5×PBS buffer, 4 μL ddH 2 Mix O evenly, react in a water bath at 95 °C for 5 min and gradually cool to room temperature to obtain 10 μM Apt-T hybrid chain solution.

Embodiment 2

[0040] Example 2 Apt-T hybrid chain concentration screening

[0041] (1) The gold electrode was polished in 0.3 µM and 0.05 µM alumina slurry in turn until it became a mirror surface, and then PBS buffer and ddH were used in turn. 2 O were washed 3-5 times repeatedly;

[0042](2) 10 μL solution of 2 μM H1 chain was added dropwise to the surface of the gold electrode obtained in step (1), incubated at 37 °C for 2 h, washed with PBS buffer 3 times to obtain H1 modified gold electrode; then 30 μM H2 chain was added 10 μL was added dropwise to the surface of the H1-modified gold electrode, incubated at 37 °C for 2 h, and washed three times with PBS buffer to obtain the H1-H2-modified gold electrode;

[0043] (3) Add 0 μL, 0.25 μL, 0.5 μL, 1 μL, 2 μL, and 4 μL of the Apt-T hybrid chain solution obtained in Example 1 to 1 μL of 10 nM ampicillin, 1 μL of 20 μM H3 chain solution, and 100 μM, respectively. 3 μL of H4 chain solution, add ddH 2 O supplemented to 10 μL, mixed evenly, a...

Embodiment 3

[0047] Example 3 Screening of the molar ratio of H1 chain and H2 chain

[0048] (1) The gold electrode was polished in 0.3 µM and 0.05 µM alumina slurry in turn until it became a mirror surface, and then PBS buffer and ddH were used in turn. 2 O were washed 3-5 times repeatedly;

[0049] (2) 10 μL solution of 2 μM H1 chain was added dropwise to the surface of the gold electrode obtained in step (1), incubated at 37 °C for 2 h, and washed with PBS buffer three times to obtain the H1 modified gold electrode; , 10 μM, 20 μM, 30 μM, 40 μM, and 50 μM of H2 chain 10 μL were added dropwise to the surface of the H1-modified gold electrode, incubated at 37 °C for 2 h, rinsed three times with PBS buffer to obtain H1-H2 modified gold electrode;

[0050] (3) Add 1 μL of Apt-T hybrid chain solution obtained in Example 1, 1 μL of 10 nM ampicillin, 1 μL of 20 μM H3 chain solution and 5 μL of 100 μM H4 chain solution, and add ddH 2 Add O to 10 μL, mix well, and react in a water bath at 37 ...

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Abstract

The invention provides an electrochemical aptamer sensor for ampicillin detection, comprising a detection electrode, heme, H3 chain, H4 chain, aptamer Apt, T chain; the detection electrode is a gold electrode with surface modified H1 chain and H2 chain, the H1 chain and H2 chain sequence are shown in SEQ ID NO: 1-2, the 5' end of the H1 chain and the H2 chain The 3' end of the modified sulfhydryl group. The sensor realizes the increase of the signal through the circulation of the T chain and the DNA walker on the electrode, which greatly improves the detection sensitivity and obtains a lower detection limit, which can be used to detect the trace amount of ampicillin in food.

Description

technical field [0001] The invention belongs to the technical field of biosensors, and relates to an electrochemical aptamer sensor for ampicillin detection. Background technique [0002] Ampicillin (AMP) is a beta-lactam antibiotic that belongs to the penicillin class and is the most commonly used class of antibiotics. Ampicillin inhibits bacterial wall synthesis and is widely used in human and veterinary treatments to prevent and treat bacterial infections of the gastrointestinal tract, respiratory tract, skin, and urogenital tract caused by bacterial infections such as Streptococcus and Staphylococcus. However, excessive use of ampicillin or ampicillin residues in food or the environment can lead to serious food safety and environmental safety problems, causing allergic reactions and symptoms such as epilepsy in humans. It is of great significance to establish efficient and reliable detection methods, such as high performance liquid chromatography (HPLC), enzyme-linked i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/327G01N27/48G01N33/53
CPCG01N27/3276G01N27/3277G01N27/48G01N33/5308G01N2415/00
Inventor 刘素江龙王玉黄加栋李莎莎孙文玉王业茹张曼茹
Owner UNIV OF JINAN
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