Unmarked aptamer fluorescence sensor for detecting tetracycline

A nucleic acid aptamer and fluorescent sensor technology, applied in the field of analytical chemistry, can solve problems such as unsuitable for rapid detection, complex analysis process, and poor selectivity, and achieve good practical value, sensitive detection method, and high selectivity.

Inactive Publication Date: 2016-08-31
JILIN UNIV
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Problems solved by technology

[0006] At present, the commonly used detection methods for tetracycline are chromatographic analysis methods, such as capillary electrophoresis, high performance liquid chromatography, liquid chromatography-mass spectrometry, etc. operation, not suitable for on-site rapid detection
Another commonly used detection method is enzyme-linked immunosorbent assay, although its detection sensitivity is high, but the selectivity is poor and the cost is relatively expensive

Method used

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  • Unmarked aptamer fluorescence sensor for detecting tetracycline
  • Unmarked aptamer fluorescence sensor for detecting tetracycline
  • Unmarked aptamer fluorescence sensor for detecting tetracycline

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Embodiment Construction

[0032] (1) Determination of the structural changes before and after the binding of the nucleic acid aptamer to the target by circular dichroism

[0033] Materials / Reagents: Tetracycline aptamer and tetracycline were purchased from Sangon Bioengineering Co., Ltd. (Shanghai, China).

[0034]Method: Dilute tetracycline aptamer to 1 μM, take 100 μL as a sample group and put it into a cuvette with a light path of 1 cm; dilute tetracycline and tetracycline aptamer to 5 μg / mL and 1 μM respectively, then mix and vortex Spin for 8 minutes to fully react the tetracycline and the nucleic acid aptamer, take 100 μL of the mixed solution as a sample group and put it into a cuvette with a light path of 1 cm; take 100 μL of triple distilled water as a blank group and put it into another 1 cm cuvette. The circular dichroism chromatogram was measured with a model PMS 450 circular dichroism spectrometer under the conditions of spectral value range from 230nm to 330nm, scanning speed of 100nm / min...

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Abstract

An unmarked aptamer fluorescence sensor for detecting tetracycline is characterized in that tetracycline is detected through the combination of the special recognition effect of thiazole orange on a G-quadruplex structure aptamer and the specificity between the aptamer and a target object through the following steps of detecting the change of the structure of the aptamer not combined with the target object and the structure of the aptamer combined with the target object through a circular dichroism spectrum, observing the system reaction of tetracycline, the aptamer and thiazole orange by detecting a fluorescence spectrogram, establishing the unmarked aptamer fluorescence sensor based on fluorescent probe thiazole orange for detecting tetracycline, detecting the selectivity of the established unmarked aptamer fluorescence sensor for tetracycline by detecting the fluorescence spectrogram, and detecting tetracycline in an actual sample by means of the established unmarked aptamer fluorescence sensor. A simple, rapid and sensitive detecting method with high selectivity for tetracycline is provided, has good practical value and is convenient to popularize.

Description

technical field [0001] A non-labeled nucleic acid aptamer fluorescence sensor for detecting tetracycline belongs to the technical field of analytical chemistry. Background technique [0002] Tetracycline antibiotics, as a kind of broad-spectrum antibiotics, are widely used to control bacterial infections and play a pivotal role in the treatment of animal diseases. Among them, tetracycline has become the most commonly used class of antibiotics because of its high efficiency, broad spectrum and low cost. Tetracycline is not only widely used as a drug in the prevention and treatment of human and animal diseases, but also is widely used as a feed additive to improve feed utilization efficiency and promote animal growth. [0003] However, the chemical properties of tetracycline are relatively stable and have a certain degree of persistence, resulting in its residue in the environment. A large number of studies have confirmed that tetracycline is widely present in soil and ecolog...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6428G01N2021/6432
Inventor 孙春燕苏瑞芳许景月张悦吕婧淇
Owner JILIN UNIV
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