Preparation method and application of competitive type aptamer sensor for detecting oxytetracycline

An aptamer sensor, oxytetracycline technology, applied in the direction of instruments, measuring devices, scientific instruments, etc., can solve the problems of expensive, time-consuming and expensive, and achieve the effect of fast detection speed, high sensitivity, and simple operation

Inactive Publication Date: 2014-03-05
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these methods are usually ti

Method used

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Experimental program
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Embodiment 1

[0026] A method for preparing a competitive aptasensor for detecting oxytetracycline (OTC), the steps are as follows:

[0027] (1) Drop 20 μL of GO-PANI nanocomposite solution onto the working surface of the glassy carbon electrode and dry at room temperature;

[0028] (2) Immerse the (1) electrode in HAuCl 4 (1 mg / mL) working solution, the gold nanoparticles were reduced to (1) the electrode surface by electroreduction, the initial potential was -0.2 V, and the reduction time was 20, 40, 60, 80, 100, 120, 140, 160, 180 S, room temperature to dry.

[0029] (3) Ag / AgCl was used as the reference electrode, the Pt electrode was used as the counter electrode, the sensor platform obtained in (2) was used as the working electrode, and the K 3 [Fe(CN) 6 ] is the working bottom liquid, connected to the electrochemical workstation, and the current response is detected by cyclic voltammetry. The potential was set from -0.2 to +0.6 V and the scan rate was 50 mV / s.

Embodiment 2

[0031] A method for preparing a competitive aptasensor for detecting oxytetracycline (OTC), the steps are as follows:

[0032] (1) Drop 20 μL of GO-PANI nanocomposite solution onto the working surface of the glassy carbon electrode and dry at room temperature;

[0033] (2) Dip (1) electrodes into different concentrations (1×10 -4 , 1×10 -3 , 1×10 -2 , 1×10 -1 , 1, 10 mg / mL) of HAuCl 4 In the working solution, the gold nanoparticles were reduced to (1) the electrode surface by means of electroreduction, the initial potential was -0.2 V, the reduction time was 120 s, and they were dried at room temperature.

[0034] (3) Ag / AgCl was used as the reference electrode, the Pt electrode was used as the counter electrode, the sensor platform obtained in (2) was used as the working electrode, and the K 3 [Fe(CN) 6 ] is the working bottom liquid, connected to the electrochemical workstation, and the current response is detected by cyclic voltammetry. The potential was set from -0...

Embodiment 3

[0036] A method for preparing a competitive aptasensor for detecting oxytetracycline (OTC), the steps are as follows:

[0037] (1) Drop 20 μL of GO-PANI nanocomposite solution onto the working surface of the glassy carbon electrode and dry at room temperature;

[0038] (2) Immerse the (1) electrode in HAuCl 4 (1 mg / mL) working solution, the gold nanoparticles were reduced to (1) the electrode surface by electroreduction, the initial potential was -0.2 V, the reduction time was 120 s, and dried at room temperature.

[0039] (3) Drop 20 μL of different concentrations (1×10 -5 , 1×10 -4 , 1×10 -3 , 1×10 -2 , 1×10 -1 mg / L) OTC-BSA onto the electrode surface, after drying, drop BSA to block non-specific binding sites;

[0040](4) Drop 20 μL of the mixture of HRP-Apt and OTC to be tested onto the surface of the electrode, rinse and dry after 30 minutes;

[0041] (5) Using Ag / AgCl as the reference electrode, using the Pt electrode as the counter electrode, using the sensor p...

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Abstract

The invention provides a competitive type aptamer sensor for detecting oxytetracycline (OTC) in food. By using graphene oxide-polyaniline (GO-PANI) and horse radish peroxidase (HRP) as amplification strategies, the competitive type aptamer sensor is constructed for performing ultra-sensitive detection on the oxytetracycline (OTC). During construction, a GO-PANI film is dropwise added on the surface of a glassy carbon electrode, and a gold nano particle is deposited on the surface of the electrode through an electrochemical method. An OTC connection aptamer has high affinity and specificity to OTC, is connected with the OTC when being used as a capture probe, and is simultaneously connected with the HRP to form an HRP-Apt. A complete antigen (OTC-BSA (Bovine Serum Albumin)) as an immobilized antigen is connected with the surface of the electrode, and is combined with the HRP-Apt by competing with to-be-detected OTC which is added in later. The sensor connected with the HRP on the surface performs electrochemical measurement in a work solution containing HQ (Hydroquinone Quinol) and H2O2. The oxytetracycline (OTC) can be detected by using the change of an electrochemical signal. The competitive type aptamer sensor prepared by adopting the method is strong in selectivity, high in sensitivity, simple and rapid to operate, and suitable for detecting the oxytetracycline (OTC) in a food.

Description

technical field [0001] The invention belongs to the field of detection of nanometer materials and food antibiotics, and provides a competitive aptamer sensor for detecting oxytetracycline, which can be used for the detection of oxytetracycline in food. Background technique [0002] Tetracycline antibiotics are broad-spectrum antibiotics that have been widely used in agricultural operations and human medicine for the treatment of infectious diseases. They operate very efficiently at low concentrations and are completely metabolized from the body after only a short period of time. However, excessive use of tetracyclines may lead to severe allergies, bacterial resistance in human safety and safety concerns in food. Oxytetracycline (OTC) is one of the most frequently used members of the tetracycline family. The World Trade Organization (WTO) also stipulates the maximum residue of oxytetracycline in medicine and food. Recently reported detection methods for oxytetracycline (OT...

Claims

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Application Information

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IPC IPC(8): G01N27/327G01N27/48
Inventor 徐伟黄加栋刘素其他发明人请求不公开姓名
Owner UNIV OF JINAN
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