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Electrochemical biosensor for detecting ampicillin without enzyme and preparation method and application thereof

A technology of ampicillin and biosensors, applied in the direction of material electrochemical variables, scientific instruments, instruments, etc., can solve the problems of high cost, low specificity and sensitivity, and achieve the effects of easy portability, improved sensitivity and reduced cost

Active Publication Date: 2019-11-26
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 problem of relatively low specificity and sensitivity and high cost of the method for detecting ampicillin in the prior art, the present invention provides a nucleic acid adaptation based on target induction, which is easy to operate, high in specificity and sensitivity, and low in cost. Electrochemical biosensors for the detection of ampicillin by conformational changes and catalyzed hairpin self-assembly amplification (CHA) and strand displacement strategies

Method used

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  • Electrochemical biosensor for detecting ampicillin without enzyme and preparation method and application thereof
  • Electrochemical biosensor for detecting ampicillin without enzyme and preparation method and application thereof
  • Electrochemical biosensor for detecting ampicillin without enzyme and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1 Preparation of gold electrodes with modified DP and HP1.

[0052] a) Gold electrode polishing treatment: the gold electrode was polished to the mirror surface in 0.3 µm and 0.05 µm alumina slurry in turn, and washed with ultrapure water for 5 times;

[0053] b) Drop the HP1 and DP solution on the surface of the gold electrode treated in a), and incubate at 37°C for 2 h.

Embodiment 2

[0054] Example 2 Effects of different HP1 concentrations on the detection of ampicillin.

[0055] (1) Hybridize Aptamer and Primer into a Probe (100 nM) in PBS buffer;

[0056] (2) Mix Probe, HP2, fuel, and 10 nM ampicillin standard solution and drop onto electrode S1 modified with 10 µM DP and different HP1 (final concentrations are 0.1 μM, 0.5 μM, 1 μM, 1.5 μM, 2 μM) Incubate at 37 °C for 2 h on -S5;

[0057] (3) Add 5 μL of heme solution (10 μM) dropwise onto the gold electrode, and then incubate at 37 °C for 30 min;

[0058] (4) Rinse the electrode in step (3) with PBS buffer solution for 3 times;

[0059] (5) containing H 2 o 2 (10 μM) PBS buffer solution, with Ag / AgCl as the reference electrode and Pt electrode as the counter electrode, the current change was measured by differential pulse voltammetry, the potential was 0 to -0.6 V, the pulse width was 0.05 V, and the scan rate was 0.06 s.

[0060] Taking the HP1 concentration as the abscissa and the current value ...

Embodiment 3

[0061] Example 3 Effects of different DP concentrations on the detection of ampicillin.

[0062] (1) Hybridize Aptamer and Primer into a Probe (100 nM) in PBS buffer;

[0063] (2) Mix the standard solution of Probe, HP2, fuel and 10 nM ampicillin and drop it on the electrode modified with different concentrations of DP of 1 μM HP1 (final concentration is 1 μM, 5 μM, 10 μM, 15 μM, 20 μM) Incubate at 37 °C for 2 h;

[0064] (3) Add 5 μL of heme solution (10 μM) dropwise onto the gold electrode, and then incubate at 37 °C for 30 min;

[0065] (4) Rinse the electrode in step (3) with PBS buffer solution for 3 times;

[0066] (5) containing H 2 o 2 (10 mM) PBS buffer solution, with Ag / AgCl as the reference electrode and Pt electrode as the counter electrode, the current change was measured by differential pulse voltammetry, the potential was 0 to -0.6 V, the pulse width was 0.05 V, and the scan rate was 0.06 s.

[0067] With the DP concentration as the abscissa and the curren...

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Abstract

The invention relates to an electrochemical biosensor which detects ampicillin without enzyme based on a nucleic acid aptamer, which belongs to the technical field of electrochemical biosensors. According to the invention, ampicillin is detected based on target-induced conformational changes in the nucleic acid aptamer and catalytic hairpin self-assembly amplification (CHA) and strand replacementstrategies, which achieves multiple signal amplification, reduces detection limits, and improves sensitivity; gold electrodes are simple, compact, easy to carry, and can be used multiple times; simpleand sensitive detection of a target object is realized; the electrochemical biosensor has the advantages of simple preparation method, stable performance and good electrode repeatability; the reaction process does not require the participation of enzyme, which greatly reduces the cost; and the electrochemical biosensor is suitable for the practical application of ampicillin detection and biosensor industrialization in food safety.

Description

technical field [0001] The invention relates to an electrochemical biosensor based on nucleic acid aptamer for enzyme-free detection of ampicillin, in particular to a target-induced nucleic acid aptamer conformational change and catalytic hairpin self-assembly amplification (CHA) and strand displacement strategy An electrochemical biosensor for detecting ampicillin, a preparation method and an application thereof belong to the technical field of electrochemical biosensors. Background technique [0002] Antibiotics have the function of killing and destroying microbial cell structures, and are widely used in the preparation of various antibacterial drugs around the world, but at the same time, the residual problem caused by the abuse of antibiotics has become a problem that needs to be solved urgently worldwide. Overuse of antibiotics can spur infection-causing bacteria to become resistant. [0003] Ampicillin, also known as ampicillin, is a beta-lactam antibiotic that can tr...

Claims

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Application Information

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IPC IPC(8): G01N27/48G01N27/327
CPCG01N27/3277G01N27/48
Inventor 刘素张儒峰黄加栋王玉瞿晓南李莎莎赵一菡江龙张曼茹
Owner UNIV OF JINAN
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