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Receptor reagent for detecting novel coronavirus and application thereof

A coronavirus, a new type of technology, applied in the field of immune detection, can solve the problems of high technical complexity, delayed disease, and long detection window period

Pending Publication Date: 2021-09-14
CHEMCLIN DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Nucleic acid detection uses polymerase chain reaction (PCR) technology, which is much more complex than immunological detection methods, and nucleic acid detection is not easy to carry out in general laboratories
[0004] However, the detection window for 2019-nCoV antibodies is relatively long (after the virus infects the human body and the human body produces antibodies), it is easy to cause false negative judgments and delay the disease

Method used

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  • Receptor reagent for detecting novel coronavirus and application thereof
  • Receptor reagent for detecting novel coronavirus and application thereof
  • Receptor reagent for detecting novel coronavirus and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0084] Example 1: Preparation of acceptor microspheres

[0085] 1.1 Synthesis of polystyrene latex microspheres

[0086] Prepare a 100mL three-neck flask, add 40mmol styrene, 3mmol methacrylic acid, and 10mL water, stir for 10min, and pass N 2 30min;

[0087] 1) Weigh 0.11g of ammonium persulfate and 0.2g of sodium chloride, dissolve them in 40mL of water to prepare an aqueous solution. This aqueous solution is joined in the reaction system of step 1), continues logical N 2 30min;

[0088] 2) The temperature of the reaction system was raised to 70°C, and the reaction was carried out for 15 hours;

[0089] 3) Cool the emulsion after the reaction to room temperature, and filter it with a suitable filter cloth. The obtained emulsion was washed with deionized water for several times by centrifugation and sedimentation until the conductivity of the supernatant at the beginning of centrifugation was close to that of deionized water, then diluted with water, and preserved in t...

Embodiment 2

[0105] Embodiment 2: the preparation of kit

[0106] 2.1 Preparation of receptor reagents

[0107] 1) Antibody treatment: dialyze the SARS-CoV-2 spine S1 subunit antibody (or SARS-CoV-2 (2019-nCoV) nucleocapsid protein / N antibody), replace it with coating buffer, and measure the protein concentration;

[0108] 2) Receptor microspheres treatment: the receptor microspheres prepared in Example 1 were replaced by coating buffer through processes such as centrifugation and ultrasound;

[0109] 3) Coupling: The treated receptor microspheres and the treated 2019-nCoV antibody 1 are mixed and reacted. After reduction and blocking, the receptor microspheres-2019-nCoV antibody 1 is obtained, and the preservation solution is used for determination. Contain and store to obtain the receptor reagent.

[0110] 2.2 Preparation of capture reagents

[0111] 1) Antibody treatment: dialyze the 2019-nCoV spike antibody (or 2019-nCoV nucleocapsid protein / N antibody), replace it with labeling bu...

Embodiment 3

[0115] Embodiment 3: Utilize anthrone method to detect sugar content

[0116] 3.1 Microsphere sample pretreatment:

[0117] The receptor reagents containing 1 mg of receptor microspheres in Example 2 were centrifuged at 20,000 G for 40 min, the supernatant was poured off, and ultrasonically dispersed with purified water. After repeated centrifugation and dispersion three times, each was dilute to 1 mg / mL with purified water as the preparation Check samples.

[0118] 3.2 Preparation of glucose standard solution:

[0119] Prepare 1 mg / mL glucose stock solution with purified water to prepare standard solution curves of 0 mg / mL, 0.025 mg / mL, 0.05 mg / mL, 0.075 mg / mL, 0.10 mg / mL, and 0.15 mg / mL.

[0120] 3.3 Preparation of anthrone solution: Prepare 2 mg / mL with 80% sulfuric acid solution.

[0121] 3.4 Add 0.1mL glucose standard solution of each concentration and the sample to be tested to the centrifuge tube, and add 1mL anthrone test solution to each tube.

[0122] 3.5 Incubat...

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Abstract

The invention relates to a receptor reagent for detecting novel coronavirus and application thereof. The receptor comprises receptor microspheres capable of reacting with active oxygen to generate detectable chemiluminescence signals; the receptor microsphere is filled with a chemiluminescence agent, and the surface of the receptor microsphere is connected with a novel coronavirus antibody 1. The kit containing the receptor reagent is used for detecting the novel coronavirus by using a double-antibody sandwich method, the window period can be effectively shortened, the detection result is quick, and the result is reported within 30 minutes; the detection flux is large, can reach 200-500 test / hour, is suitable for large-scale sample detection, and can reduce the pollution of high-risk viruses at the same time.

Description

technical field [0001] The invention belongs to the technical field of immunoassay, and in particular relates to a receptor reagent for detecting novel coronavirus and its application. Background technique [0002] Coronaviruses are a group of enveloped positive-sense single-stranded RNA viruses belonging to Nidovirales, Coronaviridae, and Coronaviridae. There are 26 known species, which are divided into Four genera (α, β, γ, and δ), of which only the α- and β-genuses contain strains that are pathogenic to humans. For a long time, coronaviruses have been important animal pathogens that can cause respiratory and intestinal diseases in mammals and birds. Among the known coronaviruses, 6 can cause human diseases, including: HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1, SARS-CoV and MERS-CoV. Among them, the first four are local epidemic diseases, mainly causing mild self-limiting diseases, while the latter two can cause severe diseases. SARS-CoV and MERS-CoV discovered in 2002...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/546G01N21/76G01N21/31
CPCG01N33/56983G01N2333/165G01N33/532G01N33/54313Y02A50/30
Inventor 吴晨刘宇卉黄正铭李临
Owner CHEMCLIN DIAGNOSTICS CO LTD
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