GII.4 norovirus detection, synthesis method thereof, and GII.4 norovirus detection method

A synthesis method and virus technology, applied in the analysis of materials, material analysis by electromagnetic means, measurement devices, etc., can solve problems such as differences in human subjective factors, and achieve stable probe binding, excellent conductivity, and good effects.

Active Publication Date: 2021-05-07
YUNNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, because it involves observation, there may be differences in human subjective factors

Method used

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  • GII.4 norovirus detection, synthesis method thereof, and GII.4 norovirus detection method
  • GII.4 norovirus detection, synthesis method thereof, and GII.4 norovirus detection method
  • GII.4 norovirus detection, synthesis method thereof, and GII.4 norovirus detection method

Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0067] Experimental Example 1: Preparation and Characterization of Fe 3 o 4 The method of @COF(TpDA) is as follows:

[0068] (1) Fe 3 o 4 Preparation of nanoclusters: Weigh ferric chloride hexahydrate (1.35g, 3mmol), ammonium acetate (3.85g, 50mmol), sodium citrate (0.4g, 1.36mmol), add 70mL ethylene glycol after mixing, Dissolve under stirring conditions, the mixture is completely dissolved to obtain a dark yellow solution, transfer the solution to a round bottom flask, continue to stir at 160 ° C for 1 h to form a uniform black solution, and then transfer the solution to a stainless steel autoclave (capacity 100mL), reacted in an oven at 200°C for 16h, after the reaction was terminated, the reaction product was naturally cooled at room temperature to obtain a suspension containing black precipitate, and the suspension was centrifuged at 8000r / min for 20min, The supernatant was removed, the precipitate was washed three times with deionized water and absolute ethanol, and ...

experiment example 2

[0070] Experimental Example 2: Preparation and Characterization of AuNPs@Fe 3 o 4 @COF(TpDA) nanocomposite approach:

[0071] (1) Preparation of AuNPs: Under stirring conditions, 1.07 mL of 23.46 mmol / L HAuCl 4 Add the solution into 100mL boiling deionized water, heat and stir at 110°C for 5min, slowly add 10mL sodium citrate solution with a concentration of 14.53mmol / L dropwise, condense and reflux for 30min until the solution turns wine red, and stir the solution at room temperature 15min, store at 4°C.

[0072] (2) Get Fe from the previous step 3 o 4 Add water to @COF to prepare Fe at a concentration of 1mg / mL 3 o 4 @COF solution, take 1mLFe 3 o 4 Add the @COF solution dropwise to 10mL gold nanoparticle solution with a concentration of 0.0002mol / L, stir at 4°C for 12h, centrifuge at 8000r / min for 10min, discard the supernatant, and continue washing with deionized water for 3 times to obtain AuNPs @Fe 3 o 4 @COF nanocomposites.

[0073] (3) Using transmission ele...

experiment example 3

[0074] Experimental Example 3: Synthesis and Characterization of WP5@AuNPs@Fe 3 o 4 @COF(TpDA) nanocomposite approach:

[0075] (1) WP5@AuNPs@Fe 3 o 4 Synthesis of @COF(TpDA) nanocomposite: Weigh 1mg WP5 powder, add 2mL deionized water to prepare 0.5mg / mL WP5 aqueous solution, slowly add 1mL WP5 aqueous solution to 2mL1mg / mL AuNPs@Fe 3 o 4 In the @COF(TpDA) solution, stir at 4°C for 4h. After the reaction, centrifuge at 8000r / min for 15min, remove the supernatant, add deionized water, repeat the centrifugation and wash three times, and put the product at -50 Freeze-dry at ℃ for 12h.

[0076] (2) Infrared spectrum, Zeta potential method is used to characterize the product, in infrared spectrum (A), 3430cm -1 and 1566cm -1 The peaks are the water and hydroxyl peaks adsorbed on the surface of the material, compared with COF (TpDA), at 571cm -1 The peak that appears at the Fe 3 o 4 The characteristic peaks of Fe-O functional groups in @COF(TpDA) are similar to those of F...

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Abstract

The invention relates to a GII.4 norovirus detection material, a synthesis method thereof, and a GII.4 norovirus detection method. The material comprises:Fe3O4@COF(TpDA) or AuNPs@Fe3O4@COF(TpDA) or WP5@AuNPs@Fe3O4@COF(TpDA), wherein the three materials are synthesized in sequence. an analyte which is a water sample, a food sample, a vomitus or an excrement sample; and an electrochemical immunosensor manufactured using the method of the invention. The linear detection range of the sensor prepared by the invention is 2.51 copies / mL to 105.4 copies / mL, and the lowest detection limit is 0.84 copy / mL.

Description

technical field [0001] The invention belongs to the field of electrochemical immunosensing, and in particular relates to a method for detecting human norovirus. Background technique [0002] Human norovirus (HuNOV) is a major cause of viral gastroenteritis worldwide. Associated with 95% of nonbacterial and 50% of gastroenteritis outbreaks worldwide. NoVs are divided into seven genogroups (GI-GVII), and each genotype is further divided into several genotypes. Human pathogens are restricted to genotypes GI, GII, and GIV, with the GII.4 subtype being the most common norovirus that infects humans, accounting for 80% of norovirus infections worldwide. [0003] Traditional detection methods include Real-time RT-PCR and ELISA. Real-time RT-PCR extracts viral RNA, designs and synthesizes primers and probes, and performs fluorescence quantification with a quantitative PCR instrument; ELISA is enzyme-linked immunosorbent assay. , this method can directly use the antibody to capture...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327
CPCG01N27/3278
Inventor 赵卉李灿鹏谢伟张亚平
Owner YUNNAN UNIV
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