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Method for detecting norovirus

A virus and solution technology, applied in the field of norovirus detection, can solve the problems of cumbersome detection of multiple virus subtypes, high detection cost, high false negative rate, etc., and achieve the effect of good HT sealing effect, excellent conductivity, and enhanced electrical signal

Active Publication Date: 2021-07-23
YUNNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Its advantages are time saving and convenient operation, so it is widely used in clinical detection, but it also has the disadvantage of high false negative rate due to its low sensitivity
Real-time RT-PCR extracts viral RNA, designs and synthesizes primers and probes, and performs fluorescence quantification with a quantitative PCR instrument; Real-time RT-PCR method is characterized by high detection sensitivity and accurate results, but for multi-virus sub- Type detection is cumbersome, and the detection cost is high, requiring expensive equipment and professional operation

Method used

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preparation example Construction

[0078] 6. The preparation method of the working electrode.

[0079] A glassy carbon electrode with a diameter of 3 mm was covered with Al 2 o 3 The polishing powder is polished into a mirror surface, and then ultrasonically cleaned in 50% nitric acid solution, 50% absolute ethanol, and ultrapure water; the electrode surface is dried with nitrogen, and 10-20 μL of AuNPs@BP at 0.5-5.0 mg / mL is added dropwise @Ti 3 C 2 MXene nanocomposite, air dry.

[0080] 7. Screen the optimal reaction conditions for the working electrode.

[0081] (1) Prepare 0.1-1mg / mL polypeptide (NoroBP) solution, take 10μL and drop it on the surface of the treated electrode, incubate at 4°C for 0-3h, wash the electrode with 0.2mol / L pH 7.0PBS 3 times, and wait for the electrode to dry After that, in 0.1mol / L KCl, 1mmol / LK 4 [Fe(CN) 6 ] / K 3 [Fe(CN) 6 ] solution for electrochemical impedance spectroscopy (EIS), while the corresponding cyclic voltammetry (CV) verification.

[0082] (2) Prepare 0.1-...

Embodiment 1

[0093] Example 1: ZnFe 2 o 4 @COF is prepared as follows:

[0094] (1) ZnFe 2 o 4 Preparation of nanoparticles: weigh ferric chloride hexahydrate (1.0 g) and anhydrous zinc chloride (0.5 g), dissolve them in ethylene glycol (60 mL) to form a transparent solution, then add sodium acetate (5.0 g) and poly Ethylene glycol 20000 (0.1 g). 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.

[0095] (2) Further synthesis of ZnFe 2 o 4 @COF: Take 1,3,5-tris(4-aminophenyl)benzene (TAPB,0.1g), terephthalaldehyde (TPA,0.06g) and ZnFe 2 o 4 (2.0g), dissolved with 50mL dimethyl sulf...

Embodiment 2

[0096] Embodiment 2: Preparation and Characterization of ZnFe 2 o 4 @COF's as follows:

[0097] (1) ZnFe 2 o 4Preparation of nanoparticles: weigh ferric chloride hexahydrate (3.0 g) and anhydrous zinc chloride (1.0 g), dissolve them in ethylene glycol (80 mL) to form a transparent solution, then add sodium acetate (2.0 g) and poly Ethylene glycol 20000 (0.1 g). 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. ZnFe 2 o 4 Nanoparticles figure 1 TEM and figure 2 SEM diagram.

[0098] (2) Further synthesis of ZnFe 2 o 4 @COF: Take 1,3,5-tris(4-aminophenyl)benzene (T...

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Abstract

The invention relates to a method for detecting norovirus. According to the method, three substances, namely AuNPs@ZnFe2O4@COF, Apt@AuNPs@ZnFe2O4@COF and AuNPs@BP@Ti3C2MXene, are utilized. The method comprises the following steps: 1) synthesizing ZnFe2O4@COF; synthesizing AuNPs@ZnFe2O4@COF; and synthesizing apt@AuNPs@ZnFe2O4@COF; 2) stripping black phosphorus BP; synthesizing BP@Ti < 3 > C < 2 > MXene; synthesizing AuNPs@BP@Ti < 3 > C < 2 > MXene; immobilizing the polypeptide (NoroBP); and 3) constructing the norovirus electrochemical sensor. The invention constructs an electrochemical sensor capable of rapidly and effectively detecting the concentration of norovirus in a sample.

Description

technical field [0001] The invention belongs to the technical field of aptamer electrochemical sensing, in particular to the technical field of a norovirus detection method. Background technique [0002] Norovirus (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. NoV can be divided into seven genomes (GI-GVII) according to their VP protein coding sequences, among which the GI, GII and GIV genomes mainly cause human infection, the GI genome contains 9 different genotypes, and the GII genome contains 22 different genotypes. The GIV genome contains two different genotypes, among which the GII genome is the most common in human acute gastroenteritis. Worldwide, the GII.4 genotype once dominated NoV infection outbreaks. [0003] Traditional detection methods include ELISA, immunochromatography and Real-time...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/30G01N27/38G01N27/327G01N27/48
CPCG01N27/308G01N27/38G01N27/307G01N27/3277G01N27/3278G01N27/48
Inventor 赵卉李灿鹏张亚平刘会芳
Owner YUNNAN UNIV
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