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Method for colorimetric detection of kanamycin based on nano-enzyme and application thereof

A kanamycin and nanozyme technology, which is applied in the field of analysis and detection, can solve the problems that kanamycin and aptamer cannot be smoothly combined, affect the effect of the assay, and the sensitivity is not high enough, and achieves low cost, convenient operation and selection. good effect

Pending Publication Date: 2022-05-10
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this method is a signal-weakening type of detection, which is easily interfered by many factors, resulting in false positive signals
In addition, it has also been reported to use gold nanoparticles [Tarun KumarSharma, Rajesh Ramanathan, Pabudi Weerathunge, Mahsa Mohammadtaheri, Hemant KumarDaima, Ravi Shukla and Vipul Bansal.Chem.Commun., 2014,50,15856--15859] or WS2 nanorods [Yue Tang, Yang Hu, Pei Zhou, Chunxiao Wang, Han Tao, and YuangenWu.J.Agric.Food Chem.2021,69,2884-2893] as a nanozyme, using the combination of kanamycin and nucleic acid aptamer to induce nucleic acid Dissociation of aptamers on the surface of nanozymes to affect the activity of nanozymes for the determination of kanamycin
However, these methods are not sensitive enough due to the lack of signal amplification reactions other than the nanozyme reaction
Moreover, due to the possible irreversible binding of the nucleic acid aptamer on the surface of the nanozyme, the target kanamycin and the aptamer cannot be smoothly combined, thereby affecting the effect of the assay

Method used

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  • Method for colorimetric detection of kanamycin based on nano-enzyme and application thereof
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  • Method for colorimetric detection of kanamycin based on nano-enzyme and application thereof

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Embodiment 1

[0039] A method for the colorimetric detection of kanamycin based on nanozyme, comprising the following steps:

[0040] (1) CuWO 4 Synthesis of nanomaterials: 0.34g Cu(NO 3 ) 2 ·3H 2 O was dissolved in 50 mL of deionized water. Then, the pH of the solution was adjusted to 5.0 with 2.0 M NaOH solution, and stirred at 60 °C for 30 min. Next, add 0.60gNa 2 WO 4 2H 2 O with continuous stirring at 70 °C for 40 min. After cooling to room temperature, wash with water and ethanol three times in sequence, and then dry at 60°C for 12 hours, and the obtained product was annealed at 400°C for 1 hour, and finally black CuWO was obtained. 4 powder, its scanning electron microscope picture is attached figure 2 As shown, its X-ray diffraction pattern is shown in the attached image 3 shown.

[0041] Depend on figure 2 It can be seen from the scanning electron microscope that the synthesized CuWO 4 Nanomaterials have irregular morphology with a size of 50-100nm. image 3 Show a...

Embodiment 2

[0049] A method for the colorimetric detection of kanamycin based on nanozyme, comprising the following steps:

[0050] (1) CuWO 4 Synthesis of nanomaterials: 0.34g Cu(NO 3 ) 2 ·3H 2 O was dissolved in 50 mL of deionized water. Then, the pH of the solution was adjusted to 5.0 with 2.0 M NaOH solution, and stirred at 60 °C for 50 min. Next, add 0.60gNa 2 WO 4 2H 2 O with continuous stirring at 80 °C for 50 min. After cooling to room temperature, wash with water and ethanol three times in sequence, and then dry at 60°C for 12 hours, the obtained product was annealed at 500°C for 1 hour, and finally black CuWO was obtained. 4 powder.

[0051] (2) Biological reaction: firstly, mesoporous silica (MSN) was prepared, and 0.3 g of cetyltrimethylammonium bromide (CTAB) was dissolved in 150 mL of deionized water. Then, 1.0 mL of 2.0 M NaOH solution was added to the above solution and stirred at 50 °C for 30 min. Subsequently, 1.5 mL of tetraethylorthosilicate (TEOS) was slowl...

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Abstract

The invention discloses a method for colorimetric detection of kanamycin based on nano-enzyme and application of the method. The method comprises the following steps: coating potassium ferricyanide in mesoporous silica, and adsorbing a nucleic acid aptamer on the surface of the mesoporous silica as biological gating; the specific recognition reaction of the target object kanamycin and the nucleic acid aptamer causes the gate control of the nucleic acid aptamer to be separated from the surface of the mesoporous silica, and causes the potassium ferricyanide to be released into a solution from the interior of the mesoporous silica, so that the surfaces of the CuWO4 nanoparticles are combined to generate nano-enzyme to induce a chromogenic reaction; during the period, the aptamer in the combination product of the kanamycin and the aptamer is sheared through deoxyribonuclease I, the kanamycin is released into the solution again, a new round of target object / aptamer combination reaction and release of potassium ferricyanide are induced again, and a detection signal is amplified; when the method is used for detecting the kanamycin, biomolecular marking and immobilization are not needed, the operation is simple and convenient, the cost is low, and the selectivity is good.

Description

technical field [0001] The invention relates to the technical field of analysis and detection, in particular to a nanozyme-based colorimetric detection method for kanamycin and its application. Background technique [0002] Kanamycin is an important antibiotic used in the treatment and prevention of microbial infections, but excessive use can lead to a series of adverse reactions, including ear and kidney toxicity and anaphylactic shock. The European Union has stipulated the maximum residues of kanamycin in food. Therefore, the detection of kanamycin is of great significance for food safety supervision. At present, kanamycin mainly relies on microbial methods, chromatography and enzyme-linked immunoassays for detection. However, these methods often require expensive instruments and equipment, time-consuming operation, require professional personnel to operate, and are not suitable for screening of large batches of samples. In contrast, the colorimetric method has the adva...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/78
CPCG01N21/78Y02A50/30
Inventor 王光丽陈彦如赵玲玲张岚
Owner JIANGNAN UNIV
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