Renewable electrochemical sensor for detecting trace kanamycin, and preparation method and application thereof

A kanamycin and biosensor technology, applied in the field of biosensors, can solve the problems of low specificity, sensitivity, and high cost, and achieve the effects of high specificity, low detection limit, and fast detection speed

Active Publication Date: 2020-07-31
SHANDONG UNIV QILU HOSPITAL +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In order to solve the problems of low specificity and sensitivity and high cost of the method for detecting trace Kana in water samples in the prior art, the present invention pr

Method used

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  • Renewable electrochemical sensor for detecting trace kanamycin, and preparation method and application thereof
  • Renewable electrochemical sensor for detecting trace kanamycin, and preparation method and application thereof
  • Renewable electrochemical sensor for detecting trace kanamycin, and preparation method and application thereof

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Experimental program
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Effect test

Embodiment 1

[0063] A preparation method of the electrochemical biosensor of the present invention:

[0064] 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;

[0065] b. Add 10 μL of 10 μM H3 dropwise to the electrode surface, incubate at 37 °C for 2 h, and wash.

[0066] 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:

[0067] a. Add 200 μL 0.5 μM magnetic beads-CP (based on CP) and 200 μL 0.5 μM H1 into 4.6 mL water sample to be tested (Kana concentration is 100 pM), shake for 30 s, and place in a constant temperature of 37 ℃ Incubate in the box for 30 min; then, use a magnet to remove the supernatant through magnetic separation to obtain magnetic beads-CP / H1 / Kana, which are dispersed in 20 μL of ultrapure water;

[0068] b. Take 20 μL ma...

Embodiment 2

[0075] A preparation method of the electrochemical biosensor of the present invention:

[0076] 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;

[0077] b. Add 10 μL of 10 μM H3 dropwise to the electrode surface, incubate at 37 °C for 2 h, and wash.

[0078] 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:

[0079] a. Add 200 μL 0.5 μM magnetic beads-CP (based on CP), 200 μL 0.5 μM H1 to 4.6 mL water sample to be tested (Kana concentration: 0, 1 fM, 5 fM, 50 fM, 500 fM, 5 pM, 50 pM, 100 pM, 500 pM), shaken for 30 s, and incubated in a 37 °C incubator for 30 min; then, the supernatant was removed by magnetic separation with a magnet to obtain magnetic beads-CP / H1 / Kana, dispersed in 20 μL ultrapure water;

[0080] b. Take 20 μL...

Embodiment 3

[0086] A preparation method of the electrochemical biosensor of the present invention:

[0087] 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;

[0088] b. Add 10 μL of 10 μM H3 dropwise to the electrode surface, incubate at 37 °C for 2 h, and wash.

[0089] 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:

[0090] e. Add 200 μL of 0.5 μM magnetic beads-CP (based on CP) and 200 μL of 0.5 μM H1 into 4.6 mL of the water sample to be tested (Kana concentration is 100 pM), shake for 30 s, and place in a constant temperature of 37 °C Incubate in the box for 30 min; then, use a magnet to remove the supernatant through magnetic separation to obtain magnetic beads-CP / H1 / Kana, which are dispersed in 20 μL of ultrapure water;

[0091] f....

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Abstract

The invention relates to the technical field of biosensors, and especially relates to a renewable electrochemical sensor for detecting trace kanamycin, and a preparation method and an application thereof. The invention aims to solve the problems of low specificity and sensitivity and high cost of the Kana detection method in the prior art. The renewable electrochemical sensor for detecting trace kanamycin is characterized in that an electrode is sequentially modified with an H3 layer, a magnetic bead-CP/H1/H2 layer and an FP layer. The preparation method comprises the following steps: pretreating the electrode; modifying the surface of the electrode with the H3 layer; extracting Kana in a water sample by using magnetic beads, and modifying the surface of the electrode with the magnetic bead-CP/H1/H2 layer; and modifying the surface of the electrode with the FP layer. The specific recognition of an aptamer by Kana is utilized; by utilizing the magnetic beads, the extraction of trace Kana is realized; an Nb.BbvCI enzyme is utilized to realize the effect of signal amplification; and the electrode regeneration process is realized by using FP.

Description

technical field [0001] The invention relates to the technical field of biosensors, in particular to a reproducible electrochemical sensor for detecting trace amounts of kanamycin and its preparation method and application. Background technique [0002] Kanamycin (Kana) is a class of aminoglycoside antibiotics produced by Streptomyces fermentation, which can be enriched in the human body through the food chain, and has potential dangers to human health, such as kidney toxicity and allergic reactions. The commonly used methods for detecting trace Kana in water samples include capillary electrophoresis, high performance liquid chromatography, and fluorescence resonance. These methods often have problems such as complicated instrument operation and high detection cost. Therefore, it is urgent to construct an analytical method for the effective detection of trace Kana in water samples. Contents of the invention [0003] In order to solve the problems of low specificity and se...

Claims

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Application Information

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IPC IPC(8): G01N27/327G01N27/48
CPCG01N27/3275G01N27/48
Inventor 刘倩宋晓蕾刘素王玉黄加栋
Owner SHANDONG UNIV QILU HOSPITAL
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