Detection method and detection kit for SARS-CoV-2 neutralizing antibody

A technology of antibody detection and detection method, applied in the field of biomedical detection, can solve the problems of not being able to know the infection risk of the tested person, slow response of neutralizing antibody detection, narrow linear range, etc., to achieve high accuracy, widen linear range, and sensitivity high effect

Inactive Publication Date: 2021-02-23
NANJING LEADING BIOMEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to solve the slow response of SARS-CoV-2 neutralizing antibody detection in the prior art, high operation requirements, narrow linear range, and can only determine whether the de

Method used

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  • Detection method and detection kit for SARS-CoV-2 neutralizing antibody
  • Detection method and detection kit for SARS-CoV-2 neutralizing antibody
  • Detection method and detection kit for SARS-CoV-2 neutralizing antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1 Reagent Preparation 1

[0040] Preparation of Reagent R1

[0041] The latex microspheres coupled with the S protein receptor binding domain (RBD) of SARS-CoV-2 in R1 in the kit are polystyrene latex microspheres with a diameter of 278nm and surface carboxyl modification (category number P0113, Manufacturer JSR Life Sciences Corporation).

[0042] Marking: Polystyrene latex microspheres with a diameter of 278nm and surface carboxyl modification are placed in 0.1M Hepes solution (pH=7.5), and 0.1M EDC (pH 6.3) is added to the latex solution to activate at 37°C for 0.5h, stirring at a constant speed Stir and react on the instrument (300r / min) for 0.5h, then add 0.01M NHS and react for 20min, then add RBD protein for coupling reaction for 1.5h, add blocking agent after the reaction is completed for 2.5h, and finally age at 37°C for 24h.

[0043] In this embodiment, the reaction solution R1 includes RBD-labeled latex microspheres, buffer solution, sugar, salt, ...

Embodiment 2

[0052] Example 2 Reagent Preparation 2

[0053] Preparation of Reagent R1

[0054] The latex microspheres coupled with the S protein receptor binding domain (RBD) of SARS-CoV-2 in R1 in the kit are polystyrene latex microspheres with a diameter of 100 nm and surface carboxyl modification (category number P0113, Manufacturer JSR Life Sciences Corporation).

[0055] Marking: Polystyrene latex microspheres with a diameter of 100nm and surface carboxyl group modification were activated in 0.1M phosphate buffer (pH=6.8), and 0.05M EDC (pH 6.0) was added to the latex solution for 0.5h at 37°C. Stir and react on a stirrer (100r / min) for 0.5h, then add 0.01M NHS and react for 10min, then add RBD protein for coupling reaction for 1h, after the reaction is completed, add a blocking agent to seal for 2h, and finally age at 37°C for 12h.

[0056] In this embodiment, the reaction solution R1 includes RBD-labeled latex microspheres, buffer solution, sugar, salt, stabilizer (sealing agen...

Embodiment 3

[0065] Example 3 Reagent Preparation 3

[0066] Preparation of Reagent R1

[0067] The latex microspheres coupled with the S protein receptor binding domain (RBD) of SARS-CoV-2 in R1 in the kit are polystyrene latex microspheres with a diameter of 400 nm and surface carboxyl modification (category number P0113, Manufacturer JSR Life Sciences Corporation).

[0068] Marking: Polystyrene latex microspheres with a diameter of 400nm and surface carboxyl modification were activated in 0.1M MES buffer (pH=6.0), and 0.2M EDC (pH 6.5) was added to the latex solution for 0.5h at 37°C. Stir and react on a stirrer (500r / min) for 0.5h, then add 0.2M NHS and react for 30min, then add RBD protein for coupling reaction for 2h, after the reaction is completed, add a blocking agent for blocking for 3h, and finally age at 37°C for 48h.

[0069] In this embodiment, the reaction solution R1 includes RBD-labeled latex microspheres, buffer solution, sugar, salt, stabilizer (sealing agent), prese...

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Abstract

The invention discloses a detection method and a detection kit for an SARS-CoV-2 neutralizing antibody and belongs to the technical field of biomedical detection. The invention discloses a detection method of an SARS-CoV-2 neutralizing antibody. The detection method is a latex enhanced immunoturbidimetry detection method based on hACE2-RBD amplification. The detection kit comprises a first latex microsphere marked by an S protein receptor binding domain RBD of SARS-CoV-2 and a second latex microsphere marked by human hACE2. By labeling the antigen on the latex microsphere, the detection signalis amplified, detection sensitivity is improved, and the detection range is widened. Whether the detected person is an infected person or not can be determined, and the infection risk of the detectedperson can be obtained. The method is advantaged in that detection of the neutralizing antibody can also be used for evaluating the clinical effect after inoculation of the SARS-CoV-2 vaccine, and has great significance for research, development and inoculation of the SARS-CoV-2 vaccine.

Description

technical field [0001] The invention belongs to the technical field of biomedical detection, and in particular relates to a detection method and a detection kit for a SARS-CoV-2 neutralizing antibody. Background technique [0002] SARS-CoV-2 is a new type of coronavirus belonging to the β genus. It is round or oval in shape, with a diameter of 60-140nm and a crown-like shape under an electron microscope. It can infect mammals and humans. After SARS-CoV-2 infection, the incubation period ranges from 2 to 14 days, and in some cases the incubation period even reaches 21 days. The initial infection ranges from asymptomatic to some symptoms such as fever, dry cough, upper respiratory tract infection, loss of appetite, and diarrhea. , Some cases gradually developed into respiratory distress, pneumonia, pulmonary hyaline lesions, hemoptysis, etc. Severe cases were accompanied by lymphopenia, C-reactive protein and lactate dehydrogenase increased. The research results show that it ...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/569G01N33/543
CPCG01N33/54313G01N33/56983G01N33/6854G01N2333/165G01N2469/20
Inventor 张玉基叶生宝王鹏李叶徐达王维
Owner NANJING LEADING BIOMEDICAL TECH CO LTD
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