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Tetracyclines aptamer and aptamer electrochemical biological sensor for detection of tetracyclines

A tetracycline aptamer and biosensor technology, applied in the field of detection, can solve the problems of long detection time, high cost of tetracycline detection, complex detection scheme, etc., and achieve low detection cost, convenient and fast modification, and no immunogenicity.

Active Publication Date: 2013-10-23
JINAN UNIVERSITY
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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 the existing tetracycline detection cost, complex detection scheme, and long detection time, the first purpose of the present invention is to provide a tetracycline aptamer

Method used

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  • Tetracyclines aptamer and aptamer electrochemical biological sensor for detection of tetracyclines
  • Tetracyclines aptamer and aptamer electrochemical biological sensor for detection of tetracyclines
  • Tetracyclines aptamer and aptamer electrochemical biological sensor for detection of tetracyclines

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: Construction of an aptamer electrochemical biosensor

[0037] (1) Polishing and activation treatment of bare gold electrode surface

[0038] Grind the bare gold electrode with 0.3 μm alumina powder for 10 minutes, then polish it with 0.05 μm alumina powder for 20 minutes, and then ultrasonically clean it twice with ultrapure water for 5 minutes each time to completely remove the non-specific adsorption on the bare gold electrode. Aluminum oxide powder on the electrode surface. Immerse the cleaned bare gold electrode in freshly prepared hot Piranha solution (concentrated sulfuric acid: 30% H 2 o 2 =7:3, V:V) were activated at room temperature for 10 minutes to obtain activated bare gold electrodes. Then ultrasonic cleaning was performed twice with ultrapure water and absolute ethanol, 5 minutes each time.

[0039] (2) Modification of aptamer electrochemical sensors

[0040] Take 2 μL of tetracycline aptamer solution with a concentration of 100 μmol / L and ...

Embodiment 2

[0043] Example 2: Electrochemical response of tetracycline aptamer electrochemical biosensor to tetracycline standard solution

[0044] (1) The electrochemical nucleic acid aptamer electrochemical biosensor produced in Example 1 was used as the working electrode, the Ag / AgCl (saturated KCl) electrode was used as the reference electrode, and the platinum wire electrode was used as the counter electrode. The electrochemical nucleic acid aptamer electrochemical biosensor made in Example 1 was rinsed with ultrapure water to remove unbound aptamers, and then placed in a concentration of 5 mmol / L K 3 [Fe(CN) 6 ] / K 4 [Fe(CN) 6 ](1:1) In the electrolyte, the electrolyte contains KCl with a concentration of 0.1mol / L, AC impedance analysis, initial potential 0.22V, frequency: 0.1-1.0×10 5 Hz, the AC impedance spectrum and its impedance value Ret0 of the aptamer electrochemical biosensor were obtained.

[0045] (2) Rinse the aptamer electrochemical biosensor described in step (1) wit...

Embodiment 3

[0048] Example 3: Stability and reproducibility of the aptamer electrochemical biosensor

[0049] The electrode detection signal comes from the aptamer’s recognition of tetracycline. The combination of tetracycline and aptamer causes a change in the signal. The number of aptamers assembled on the bare gold directly determines the detection sensitivity of the aptamer electrochemical biosensor. In order to intuitively reflect the effect of electrode assembly and at the same time reflect the reproducibility of aptamer electrochemical biosensors, an evaluation standard --- assembly rate Q was set.

[0050] The prescribed assembly rate Q is: Q = Ip ( Au ) - Ip ( AP ) Ip ( Au ) ...

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PUM

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Abstract

The invention relates to a tetracyclines aptamer, an aptamer electrochemical biological sensor for detection of tetracyclines, and a preparation method and a detection method of the sensor. The tetracyclines aptamer is a single strand DNA probe containing a TTGAGCCCT sequence and having the length of 13-20 bp, and an amino is modified by a 5' end, namely 5'-NH2-(CH2)6-. The aptamer electrochemical biological sensor for the detection of the tetracyclines contains the tetracyclines aptamer. The sensor improves the sensitivity of the detection of the tetracyclines, and can detect the tetracyclines in a minimum concentration of 1.0 ng / mL; and the sensor greatly reduces the cost of the detection of the tetracyclines, has simple and convenient operation, and can rapidly detecting the tetracyclines.

Description

technical field [0001] The invention belongs to the technical field of detection, and relates to a tetracycline aptamer, an aptamer electrochemical biosensor for detecting tetracycline, and a preparation method and a detection method of the sensor. Background technique [0002] Tetracyclines (tetracyclines, TCs) are a class of broad-spectrum antibacterial drugs containing a parallel tetraphenyl structure produced by actinomycetes. Both have inhibitory effects, and their antibacterial mechanism is mainly to inhibit bacterial protein synthesis. At present, tetracycline antibiotics have become the antibiotics with the largest production and clinical use in my country's livestock and poultry industry. Tetracycline can be directly put into the feed as a feed additive, and can also be used as a growth promoter to feed animals, occupying an important position in the pharmaceutical market. However, the abuse of tetracyclines brings great harm to the health of people and animals, a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/115G01N27/327G01N27/416
Inventor 刘大岭陈丹谢春芳姚冬生
Owner JINAN UNIVERSITY
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