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Magnetic nano-enzyme material with peroxidase catalytic activity and kit for detecting norovirus and application thereof

A technology of peroxidase and magnetic nanometers, applied in metal/metal oxide/metal hydroxide catalysts, organic compound/hydride/coordination complex catalysts, physical/chemical process catalysts, etc., can solve the problem of natural enzymes HRP is unstable, variable and inactivated, and ELISA detection is inconvenient for popularization and application, etc., to achieve good selectivity, good anti-interference, and high use value.

Active Publication Date: 2021-09-03
YUNNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the instability of the natural enzyme HRP under non-physiological conditions, it is easy to denature and inactivate, which brings inconvenience to the popularization and application of traditional ELISA detection.

Method used

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  • Magnetic nano-enzyme material with peroxidase catalytic activity and kit for detecting norovirus and application thereof
  • Magnetic nano-enzyme material with peroxidase catalytic activity and kit for detecting norovirus and application thereof
  • Magnetic nano-enzyme material with peroxidase catalytic activity and kit for detecting norovirus and application thereof

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preparation example Construction

[0040] In the present invention said ZnFe 2 o 4 The preparation method of @COF preferably includes the following steps:

[0041] 1) Combine 1,3,5-tris(4-aminophenyl)benzene, terephthalaldehyde and ZnFe 2 o 4 Dissolved in dimethyl sulfoxide to obtain a premixed solution;

[0042] 2) Under the action of anhydrous acetic acid, the premixed solution is subjected to a compound reaction, and the reactant obtained is ZnFe 2 o 4 @COF.

[0043] In the present invention, the 1,3,5-tri(4-aminophenyl)benzene, terephthalaldehyde and ZnFe 2 o 4 The mass ratio is preferably 0.1-0.8:0.01-0.2:0.1-1.0, more preferably 0.2-0.6:0.05-0.15:0.4-0.8, most preferably 0.4:0.1:0.6. After dissolving with dimethyl sulfoxide, the concentration of 1,3,5-tris(4-aminophenyl)benzene is preferably 0.002-0.016g / ml, more preferably 0.004-0.012g / ml, most preferably 0.008g / ml. In the present invention, there is no special limitation on the sources of the 1,3,5-tris(4-aminophenyl)benzene, terephthalaldehy...

Embodiment 1

[0078] ZnFe 2 o 4 @COF is prepared as follows:

[0079] (1) ZnFe 2 o 4 Preparation of nanoparticles: 1.0 g of ferric chloride hexahydrate and 0.5 g of anhydrous zinc chloride were weighed, dissolved in 60 mL of ethylene glycol to form a transparent solution, and then 5.0 g of sodium acetate and 0.1 g of polyethylene glycol 20000 were added. After the mixture was vigorously stirred for 2 h, it was transferred to a stainless steel autoclave (capacity 100 mL) and reacted at 200 ° C for 12 h. After the reaction was terminated, it was naturally cooled to room temperature to obtain a suspension containing black precipitate. The precipitate was washed several times with ethanol and water, and the black product was dried at 60° C. under vacuum for 6 h. Obtain powdered ZnFe 2 o 4 nanoparticles.

[0080] (2) Synthesis of ZnFe 2 o 4 @COF: Take 0.1g 1,3,5-tris(4-aminophenyl)benzene (TAPB), 0.06g terephthalaldehyde (TPA) and 2.0g ZnFe 2 o 4 , dissolved with 50 mL dimethyl sulfox...

Embodiment 2

[0082] Preparation and Characterization of ZnFe 2 o 4 @COF

[0083] (1) ZnFe 2 o 4 Preparation of nanoparticles: Weigh 3.0g ferric chloride hexahydrate and 1.0g anhydrous zinc chloride, dissolve in 80mL ethylene glycol to form a transparent solution, then add 2.0g sodium acetate and polyethylene glycol 20000 (0.1g) . After the mixture was vigorously stirred for 0.5 h, it was transferred to a stainless steel autoclave (capacity 100 mL) and reacted at 200 ° C for 24 h. After the reaction was terminated, it was naturally cooled to room temperature to obtain a suspension containing black precipitate. The precipitate was washed several times with ethanol and water, and the black product was dried at 60° C. under vacuum for 6 h. Obtain powdered ZnFe 2 o 4 nanoparticles. The prepared ZnFe 2 o 4 The nanoparticles were tested by TEM and SEM respectively, and the test results are shown in image 3 A and image 3 b.

[0084] (2) Synthesis of ZnFe 2 o 4 @COF: Take 0.3g 1,3,...

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Abstract

The invention provides a magnetic nano-enzyme material with peroxidase catalytic activity and a kit for detecting norovirus and an application thereof, and belongs to the technical field of virus detection. The magnetic nano-enzyme material with peroxidase catalytic activity is prepared from ZnFe2O4@ COF and gold nanoparticle raw materials. The invention also provides a magnetic nano-composite enzyme for specifically detecting the norovirus. The magnetic nano-composite enzyme is formed by combining a specific norovirus recognition biological material on gold nanoparticles of the magnetic nano-enzyme material, can specifically recognize and combine the norovirus, and has efficient horseradish peroxidase catalytic activity; and a horseradish peroxidase-labeled secondary antibody in traditional ELISA detection can be completely replaced, and detection of norovirus under non-physiological conditions is realized. The detection kit provided by the invention has the advantages of convenience in detection operation, low cost, high detection throughput and higher use value.

Description

technical field [0001] The invention belongs to the technical field of virus detection, and in particular relates to a magnetic nano-enzyme material with peroxidase catalytic activity and a kit for detecting norovirus and its application. Background technique [0002] Norovirus (NoV) is a linear single-stranded positive-sense RNA virus belonging to the Caliciviridae family. It is the main pathogen that can cause epidemics and outbreaks of acute gastroenteritis in humans after rotavirus. At present, there is no vaccine available for Norovirus. Therefore, accurate and rapid detection of NoV is of great significance for the rapid diagnosis of clinicians and better treatment for patients. [0003] At present, enzyme-linked immunosorbent assay (ELISA) is still the most commonly used norovirus detection method, which has the advantages of rapidity, high throughput and low cost. The ELISA detection method of norovirus concentration in the body that has been reported at home and ab...

Claims

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Application Information

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IPC IPC(8): B01J31/06B01J23/89G01N33/569G01N33/543
CPCB01J31/06B01J31/069B01J23/8953B01J23/002G01N33/56983G01N33/54326G01N33/54306G01N2333/08B01J35/33B01J35/40
Inventor 赵卉李灿鹏张亚平刘会芳
Owner YUNNAN UNIV
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