Method for detecting tetracycline (TET) based on colloidal gold (AuNPs) chromatography test strip of aptamer

A nucleic acid aptamer and colloidal gold test paper technology, applied in the field of analytical chemistry, can solve the problems of unsuitability for rapid detection, complex analysis process, poor selectivity, etc., and achieve the effect of preventing and controlling food safety incidents

Inactive Publication Date: 2017-05-10
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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  • Method for detecting tetracycline (TET) based on colloidal gold (AuNPs) chromatography test strip of aptamer
  • Method for detecting tetracycline (TET) based on colloidal gold (AuNPs) chromatography test strip of aptamer

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Embodiment Construction

[0028] (1) Synthesis of colloidal gold nanoparticles suitable for labeling aptamers

[0029] Materials / Reagents: Auric Chloride (HAuCl 4 ·3H 2 O) (Aladdin) and trisodium citrate were purchased from Sangon Bioengineering Co., Ltd. (Shanghai, China).

[0030] METHODS: AuNPs were prepared according to a slightly modified classical Frens method. First, all glassware used for sample preparation was thoroughly cleaned with aqua regia (hydrochloric acid / HNO3, volume ratio 3:1) and dried for later use. 100 mL of 0.01% chloroauric acid solution was boiled thoroughly, and 1.8 mL of 1% trisodium citrate solution was quickly added with constant stirring. When the solution was boiled for another 5 min, the color of the solution changed from blue to wine red within 1 min. Visual inspection of the prepared colloidal gold solution showed a bright wine red color, uniform distribution, and no obvious particle sediment or floating matter. After cooling at room temperature with stirring, the...

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Abstract

The invention discloses a method for detecting tetracycline (TET) based on a colloidal gold (AuNPs) chromatography test strip of aptamer. The method uses specific binding between colloidal gold and the aptamer and competitive binding capacity strength between the tetracycline and DNA (deoxyribonucleic acid) and the aptamer. The method comprises the following steps of synthesizing colloidal gold nanoparticles suitable for marking the aptamer; assembling and coupling the colloidal gold and the aptamer; assembling streptavidin with a DNA chain; preprocessing a sample pad and a gold pad; scribing lines on an NC membrane; assembling the test strip; detecting standard tetracycline; performing selectivity and specificity experiments; using the established colloidal gold test strip based on the aptamer to measure the tetracycline in a practical sample. The tetracycline detecting method provided by the invention has the advantages of having better selectivity and flexibility, being simple, economic and easy to operate and being more suitable for in-site quick detection.

Description

technical field [0001] The invention relates to the detection of tetracycline (TET) by colloidal gold (AuNPs) chromatographic test strips based on nucleic acid aptamer (aptamer), belonging 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 s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N33/532
CPCG01N33/53G01N33/532
Inventor 孙春燕李松函李志红李锐涛李晨张子扬
Owner JILIN UNIV
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