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Aptamer electrode for detecting terramycin, and manufacturing method thereof

An oxytetracycline and electrode technology, which is applied in the preparation of electrochemical aptamer electrodes and the field of aptamer electrodes for detection of oxytetracycline, can solve the problems of complicated instrument operation and high detection cost, and achieve good repeatability, low process cost, The effect of simple preparation method

Inactive Publication Date: 2015-07-15
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the shortcomings of high detection cost in the existing detection methods in the above prior art, complicated instrument operation, and the need for professional operators, the present invention has designed a novel sandwich configuration based on nucleic acid aptamers for oxytetracycline detection. Electrochemical aptamer electrode

Method used

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Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0057] (1) Preparation of graphene oxide (GO):

[0058] Graphene oxide (GO) was prepared according to classical methods. Briefly, 1.0 g of graphite powder was added to 0.5 g of sodium nitrate (NaNO 3 ) and 3.0 g potassium permanganate (KMnO 4 ) with a mass fraction of 98% in 50 mL of concentrated sulfuric acid (H 2 SO 4 )middle. When the temperature is lower than 20 °C, react for 1 h, then raise the temperature to 35 °C, and continue to stir for 30 min. Then slowly add about 100 mL of ionized water, and after stirring continuously for 20 min, add 50 mL of H 2 o 2 (30%), to reduce the residual oxidant, the solution gradually turned bright yellow; filtered while hot, washed with HCl solution and deionized water until no sulfate was detected in the filtrate. The precipitate was dried in a thermostat at 60 °C to obtain GO.

[0059] (2) Synthesis of graphene-gold nanocomposites (rGO-Au) nanocomposites:

[0060] Glassy carbon electrodes are first polished in 0.3 and 0.05 μm...

Embodiment 1

[0075] The electrode modification process mainly includes the following steps:

[0076] a. The glassy carbon 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; the glassy carbon electrode is first polished in 0.3 and 0.05 μm alumina slurry , until it was a mirror surface, washed repeatedly with PBS and secondary water. Afterwards, the electrodes were immersed in 10 μL of the prepared graphene oxide-gold nanoparticles solution for 3 h.

[0077] b. After washing several times with secondary water, 10 μL of commercially available BSA-OTC conjugate (10 μM) solution was added dropwise on the surface of the electrode, and kept at room temperature for 150 min to completely immobilize the BSA-OTC complex on the electrode. That is, the electrode modification is completed. The thickness of the BSA-OTC coupler layer is 1 μm ± 0.2 μm,

[0078] The detection method is as follows:

[0...

Embodiment 2

[0086] With Ag / AgCl as the reference electrode and Pt electrode as the counter electrode, the potential was set from -0.2 to 0.6 V, the pulse width was 0.05 V, and the pulse width was scanned at 0.06 S. Differential pulse voltammetry was used to read the changes of electrical signals. Detect target OTC. The specific data are shown in the following table. When the concentration is 0.1 μM, the linear relationship is also met, indicating that the present invention can accurately detect oxytetracycline at a concentration of 0.1 μM. Other steps are with embodiment 1.

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Abstract

The invention provides an aptamer electrode for detecting terramycin. The electrode is characterized in that a glassy carbon electrode is sequentially modified with a graphene-gold nanometer compound and a BSA-OTC compound from bottom to top. A signal layer is methylene blue modified aptamer having a G tetrad structure and folded under an RCA chain amplification reaction, and the content of a target product is reflected through the amount of methylene blue. The invention also provides a manufacturing method of the aptamer electrode. The aptamer electrode has the advantages of simple manufacturing method, stable performances, and good repeatability, and is suitable for terramycin detection in the medicine safety and the industrialized practical application of biosensors.

Description

technical field [0001] The invention belongs to the technical field of detection sensors, and in particular relates to an aptamer electrode for detecting oxytetracycline prepared by adding a primer to an aptamer to trigger RCA and using an RCA cycle amplification method. At the same time, the invention also relates to a preparation method of an electrochemical aptamer electrode for detecting oxytetracycline. Background technique [0002] Oxytetracycline (OTC) is a tetracycline substance, a class of broad-spectrum antibiotics produced by Streptomyces, which belong to the derivatives of polycyclic tetracenecarboxamide core in chemical structure, and can be divided into natural Drugs and tetracycline antibiotics generally bind to the 30S subunit of the ribosome to prevent aminoacyl tRNA from binding to the ribosome to produce pharmacological effects, and play a bacteriostatic and bactericidal role. In the late 1940s and early 1950s, aureomycin, oxytetracycline and tetracycl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327
Inventor 王玉许颖黄加栋刘素徐伟王虹智郭玉娜邱婷婷崔杰
Owner UNIV OF JINAN
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