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Preparation method of aptamer sensor for detecting antibiotic residue

An aptamer sensor and antibiotic technology, which can be used in instruments, measuring devices, scientific instruments, etc., can solve the problems of difficulty in wide application, cumbersome operation, and high cost, and achieve the advantages of avoiding poisoning, simple deposition process, and increasing response current. Effect

Inactive Publication Date: 2014-06-04
SHANDONG UNIV OF TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although these analytical methods have high selectivity, they are difficult to be widely used due to cumbersome operation, long time-consuming, high cost, and the need to pre-concentrate and derivatize the sample

Method used

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  • Preparation method of aptamer sensor for detecting antibiotic residue
  • Preparation method of aptamer sensor for detecting antibiotic residue

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Embodiment 1

[0034] The present invention will be further described below in conjunction with embodiment, but the present invention is not limited by embodiment. Example 1 Preparation of aptasensor

[0035] A kind of preparation step of the aptamer sensor that detects kanamycin is as follows:

[0036] (1) Preparation of carbon nanotube-graphene nanocomposite film: take 100mg of carbon nanotubes and add HNO with a concentration of 2.6mol / L 3 Reflux at 140°C for 6 hours, filter with suction, and wash with ultrapure water until the pH is neutral. Graphene is added to the above solution, and the above solution is dispersed in N, N-dimethylformamide, and then the dispersion is dispersed by ultrasonic waves until a black suspension is obtained;

[0037] (2) Cleaning of the gold electrode: Before the gold electrode is modified, it is first immersed in a hot "piranha" solution (H 2 SO 4 :30%H 2 o 2 =3:1) Soak in 15min, wash with water, then use 0.3μm, 30nm Al 2 o 3 Polish the slurry on the...

Embodiment 2

[0044] Example 2 Optimization of aptamer usage conditions and determination of detection limit

[0045] 1. Optimization of aptamer usage conditions

[0046] The experimental conditions of the prepared aptamer sensor were optimized from four aspects: aptamer concentration, pH of test bottom solution, test temperature and specific binding time of aptamer and antibiotic. The measured pH of the optimal test solution was 7.5, the optimal concentration of the aptamer was 5 μg / mL, and the optimal time and temperature for the specific binding of the aptamer and kanamycin were 60 min and 35 °C, respectively; 2. Aptamer Detection Limit Determination

[0047] Prepare a standard solution of kanamycin from 0.01ng / mL to 200.0ng / mL, immerse the aptamer sensors prepared above into the standard solution of kanamycin at different concentrations, react specifically at room temperature for 60min, and detect the aptamer The current change before and after the specific reaction between the body a...

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Abstract

The invention discloses a preparation method of an aptamer sensor for detecting antibiotic residue, belonging to the technical field of safety test of agricultural products. The method comprises the steps of carrying out electrochemical polymerization on a gold electrode and thionine to obtain electrically polymerized thionine electrode; after that, evenly dripping the prepared carbon nano tube-graphene nano-composition onto the surface of the electrically polymerized thionine electrode; drying in the air, and then soaking the electrode into antibiotic aptamer solution; washing, drying in the air, and closing by bull serum albumin (BSA) solution to obtain the antibiotic aptamer sensor. The prepared aptamer sensor has the advantages of not needing pretreatment, being high in analysis speed, simple in an operation system, high in accuracy, high in specificity, low in cost, rapid in response and the like, and has the limit of detection reaching up to 0.8nm, thus being suitable for rapidly detecting the antibiotic residue.

Description

technical field [0001] The invention relates to a preparation method of an aptamer sensor for detecting antibiotic residues, belonging to the technical field of agricultural product safety detection. technical background [0002] Antibiotics, as a drug for treating diseases, have been widely used in the treatment of diseases in dairy cows. Whether through intramuscular injection or oral administration, the antibiotics entering the cows can be excreted through the metabolism of the cows, and milk is an important way for cows to excrete antibiotics from the body. Therefore, the problem of antibiotic residues in milk, the general expert survey believes that improper medication of lactating dairy cows and careless safety time are two important sources of antibiotic residues in milk, especially when using udder perfusion method to treat mastitis, it is easy cause antibiotic residues in milk. Milk is a nutritious product suitable for all ages. If antibiotics are contained in mil...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/26G01N27/327
Inventor 郭业民孙霞李发兰王相友刘君峰
Owner SHANDONG UNIV OF TECH
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