Aptamer-molecular imprinting fluorescent sensor having double specific recognition on kanamycin as well as preparation method and application of sensor

A fluorescent sensor, kanamycin technology, applied in chemical instruments and methods, fluorescence/phosphorescence, instruments, etc., can solve the problems of aptamer-molecularly imprinted fluorescent sensing that have not been reported, and achieve enhanced dual recognition effect, Good selectivity and high yield

Active Publication Date: 2018-07-06
佛山市三水佛水供水有限公司
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

However, aptamer-molecularly imprinted fluorescence sensing ba

Method used

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  • Aptamer-molecular imprinting fluorescent sensor having double specific recognition on kanamycin as well as preparation method and application of sensor
  • Aptamer-molecular imprinting fluorescent sensor having double specific recognition on kanamycin as well as preparation method and application of sensor
  • Aptamer-molecular imprinting fluorescent sensor having double specific recognition on kanamycin as well as preparation method and application of sensor

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preparation example Construction

[0063] like figure 1 As shown, the present invention provides a method for preparing an aptamer-molecularly imprinted fluorescent sensor with dual specific recognition for kanamycin, comprising the following steps. First, the template molecule kanamycin, sulfhydryl-modified Mycin aptamer, methacrylic acid to form a pre-polymerization complex; then, the aptamer-quantum dot complex is formed by the pre-polymerization complex, cross-linking agent, allylamine-modified CdSe quantum dots, and initiator; finally, the The template molecule kanamycin was eluted from the aptamer-quantum dot complex, and an aptamer-molecularly imprinted fluorescent sensor with dual specific recognition for kanamycin was obtained.

[0064] The present invention also provides the aptamer-molecular imprinted fluorescence sensor with dual specific recognition for kanamycin prepared by the above preparation method and its application in detecting the content of kanamycin in a sample.

[0065] Various test in...

Embodiment 1

[0069] (1) Synthesis of carboxyl-modified CdSe quantum dots (CdSe QDs)

[0070] A: Weigh 2.3g of selenium powder and 2.3g of sodium borohydride with an analytical balance, stir slowly under a nitrogen atmosphere to dissolve them in 25ml of ultrapure water and react. NaHSe precursor solution.

[0071] B: Weigh 0.228g 2.5 chromium chloride (CrCl 2 2.5H 2 (2), join in the three-neck round-bottomed flask of 250ml, add the ultrapure water of 100ml and make it dissolve, then slowly add 97.5 microliters of 3-mercaptopropionic acid while stirring, then add freshly prepared 1M sodium hydroxide dropwise The solution adjusted the pH of the solution to 11.

[0072] C: Nitrogen was passed for half an hour to remove the dissolved oxygen in the solution, and then the freshly prepared NaHSe precursor solution was quickly added to the mixture under vigorous magnetic stirring, and then refluxed at 100°C for an appropriate time to obtain orange transparent CdSe QDs solution.

[0073] D: Add...

Embodiment 2

[0086] (1) Synthesis of carboxyl-modified CdSe quantum dots (CdSe QDs)

[0087] The synthesis method of the above carboxyl-modified CdSe quantum dots (CdSe QDs) in this example is the same as that in Example 1.

[0088] (2) Synthesis of allylamine-modified CdSe quantum dots (CdSe QDs-AA)

[0089] The synthesis method of the above-mentioned allylamine-modified CdSe quantum dots (CdSe QDs-AA) in this example is the same as that in Example 1.

[0090] (3) Preparation of an aptamer-molecularly imprinted fluorescent sensor (apta-MIP-CdSe QDs) with dual specific recognition for kanamycin

[0091] 1) Preparation of pre-polymerized complex

[0092] Weigh 1 mg of kanamycin with a balance, dissolve it in 1 ml of phosphate buffer solution with a pH value of 7, and prepare a 1 mg / ml storage solution. Dissolve 2.8nmol of kanamycin, 2.8nmol of thiol-modified kanamycin aptamer and 5.6nmol of methacrylic acid (MAA) in 0.1ml of phosphate buffer, mix well and place in a 37°C water bath Cond...

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Abstract

The invention provides a preparation method for an aptamer-molecular imprinting fluorescent sensor having double specific recognition on kanamycin. The method comprises the following steps: firstly, the template molecule kanamycin, a thiol modified kanamycin aptamer and methacrylic acid form a prepolymerized complex; secondly, the prepolymerized complex, a crosslinking agent, allylamine modified CdSe quantum dots, and an initiator form an aptamer-quantum dot complex; and finally, the template molecule kanamycin in the aptamer-quantum dot complex is eluted, and therefore the aptamer-molecular imprinting fluorescent sensor having double specific recognition on the kanamycin is obtained. The invention also provides the aptamer-molecular imprinting fluorescent sensor having double specific recognition on the kanamycin prepared by the above preparation method, and an application of the sensor in detecting the content of the kanamycin in a sample. The aptamer-molecular imprinting fluorescentsensor provided by the invention has double specific recognition on the kanamycin, and very good selectivity on the kanamycin.

Description

technical field [0001] The invention belongs to the field of molecular imprinting preparation, and in particular relates to an aptamer-molecular imprinting fluorescent sensor with double specific recognition for kanamycin and its preparation method and application. Background technique [0002] Kanamycin is an amino antibiotic that plays an important role in the treatment and prevention of microbial infections, but excessive use can cause side effects such as allergic reactions, nephrotoxicity, and hearing loss. In addition, residual kanamycin in environmental water and food can also cause water and food pollution, thereby endangering human health. Most of the current kanamycin detection methods require expensive instruments and complicated operations, which are time-consuming and labor-intensive. Therefore, it is necessary to find a simple, rapid and selective method for the detection of kanamycin. [0003] As a recognition element, molecularly imprinted polymers have gai...

Claims

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Application Information

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IPC IPC(8): C08F222/38C08F220/06C08J9/26B01J20/26B01J20/30G01N21/64
CPCB01J20/268C08F222/385C08J9/26C08J2201/0424G01N21/643G01N2021/6439C08F220/06
Inventor 梁勇耿园园汤又文杨军张嘉霖魏伟平
Owner 佛山市三水佛水供水有限公司
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