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Novel coronavirus SARS-CoV-2 S protein detection method

A coronavirus and protein detection technology, applied in measurement devices, immunoassays, instruments, etc., can solve problems such as the inability to effectively detect the new coronavirus SARS-CoV-2 S protein, and achieve rapid biotin labeling and alkaline phosphatase labeling. , the effect of rapid preparation

Pending Publication Date: 2020-06-12
SICHUAN ORIENTER BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The invention provides a novel coronavirus SARS-CoV-2 S protein detection method, which solves the problem that the prior art cannot effectively detect the novel coronavirus SARS-CoV-2 S protein

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] S1. Angiotensin-converting enzyme 2ACE2 recombinant protein and biotin ester labeling reagent NHS-LC-Biotin were mixed and reacted at room temperature for 30 minutes at a ratio of 1:20 (mol / mol);

[0026] Mix biotinylated ACE2 recombinant protein with streptavidin magnetic beads at 5 μg / mg and react at room temperature for 30 minutes to prepare ACE2 recombinant protein-coated magnetic particles;

[0027] S2. Mix (N-maleimidomethyl)cyclohexane-1-carboxylic acid succinimidyl ester SMCC and alkaline phosphatase ALP at a ratio of 1:38 (w / w) for 30 minutes at room temperature to prepare activation Alkaline phosphatase ALP;

[0028] 2-IT and ACE2 recombinant protein were mixed at room temperature for 30 minutes at a ratio of 1:12 (w / w) to prepare activated ACE2 recombinant protein;

[0029] Mix the activated alkaline phosphatase and the activated ACE2 recombinant protein at a ratio of 1:1.2 (w / w) for 2 hours at room temperature to prepare alkaline phosphatase ALP-labeled ACE...

Embodiment 2

[0034] A kit for the detection method of the novel coronavirus SARS-CoV-2 S protein, the kit comprising: biotin ester-labeled ACE2 recombinant protein-coated magnetic particles, alkaline phosphatase ALP-labeled ACE2 recombinant protein and luminescent Substrate AMPPD. The preparation process of biotin ester-labeled ACE2 recombinant protein-coated magnetic particles is as follows: angiotensin-converting enzyme 2 ACE2 recombinant protein and biotin ester labeling reagent NHS-LC-Biotin are mixed at room temperature in a ratio of 1:20 (mol / mol). Uniformly react for 30 minutes, mix biotin-labeled ACE2 recombinant protein with streptavidin magnetic beads at 5 μg / mg and react at room temperature for 30 minutes to prepare ACE2 recombinant protein-coated magnetic particles; preparation of alkaline phosphatase ALP-labeled ACE2 recombinant protein The process is: mix (N-maleimidomethyl)cyclohexane-1-carboxylate succinimidyl ester SMCC and alkaline phosphatase ALP at a ratio of 1:38 (w / w)...

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Abstract

The invention discloses a novel coronavirus SARS-CoV-2 S protein detection method. SARS-CoV-2 virus depends on spike protein on the surface to be combined with angiotensin converting enzyme 2 on the surface of a cell so as to enter the cell; according to the method, raw materials commonly used in chemiluminescence immunoassay and ACE2 are easy to purchase from the market, the biotin labeling and the alkaline phosphatase labeling are rapid, the reagent for SARS-CoV-2 S protein detection can be rapidly prepared, and the positive coincidence rate, the negative coincidence rate and the total coincidence rate are high compared with the SARS-CoV-2 fluorescence quantitative PCR result.

Description

technical field [0001] The invention relates to a virus detection method, in particular to a novel coronavirus SARS-CoV-2 S protein detection method. Background technique [0002] Viral antigen detection and immunological analysis methods usually use the double antibody sandwich method. Taking the detection of SARS-CoV N protein antigen as an example, the solid-phase end is SARS-CoV N protein-specific antibody, and the labeled end is SARS-CoV N protein-specific antibody. After adding the analyte and incubating, the SARS-CoV N protein in the analyte will combine with the SARS-CoV N protein-specific antibody at the solid phase end to form an immune complex, and at the same time, the SARS-CoV N protein-specific antibody at the label end will combine with the SARS-CoV N protein-specific antibody in the sample. - Different antigenic sites on the CoV N protein bind to form a labeled immune complex, and the unbound enzyme conjugates and other substances are removed by washing. By ...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/543G01N33/535
CPCG01N33/535G01N33/54326G01N33/56983G01N2333/165G01N2333/948G01N2469/10
Inventor 文莉惠
Owner SICHUAN ORIENTER BIOLOGICAL TECH
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