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SARS-CoV-2 neutralizing antibody test strip and preparation method and kit thereof

A sars-cov-2 and test strip technology, applied in the field of SARS-CoV-2 neutralizing antibody test strips and its preparation, can solve the problems of inconvenient on-site detection

Active Publication Date: 2021-08-20
南京申基医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This invention marks the antigen on the latex microspheres. Although it amplifies the detection signal to increase the sensitivity of the detection, it is a turbidimetric method and requires the use of supporting equipment for detection. At the same time, the reagents need to be stored in an environment of 4°C, which is not convenient for detection. On-site detection of people in other scenarios

Method used

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  • SARS-CoV-2 neutralizing antibody test strip and preparation method and kit thereof
  • SARS-CoV-2 neutralizing antibody test strip and preparation method and kit thereof
  • SARS-CoV-2 neutralizing antibody test strip and preparation method and kit thereof

Examples

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preparation example Construction

[0042] Examples of Preparation of Raw Materials / Intermediates

[0043] Chicken IgY was purchased from Shanghai Yanmu Industrial Co., Ltd. (Cat. No. FK-025).

[0044] Goat anti-chicken secondary antibody was purchased from Luoyang Baitaike Biotechnology Co., Ltd., mg / mL titer (ELISA) > 1:100,000, polyacrylamide gel electrophoresis purity > 96%; storage buffer 0.01mol / L phosphate buffer, containing 0.14mol / L sodium chloride, pH 7.4; P300 preservative was purchased from SIGMA, Sinopharm USA.

[0045] Human IgG was purchased from Beijing Yita Biotechnology Co., Ltd., the product number is YT7150.

[0046] Anti-human IgG antibody was purchased from Shanghai Jianglai Biotechnology Co., Ltd., the product number is AbM59504-10-PU.

[0047] The SARS-CoV-2 S-RBD antigen was purchased from Nanjing Shenji Biotechnology Co., Ltd. The protein was constructed as a DNA sequence encoding the RBD region (Gln321-Ser591) of the spike protein of SARS-CoV-2 (2019-nCoV), and the expression host w...

preparation example 1

[0050] Preparation of colloidal gold

[0051] The preparation method of colloidal gold in the present application adopts sodium citrate reduction method to prepare colloidal nanoparticles, comprising the following steps:

[0052] (1) Take 99mL ultrapure water and 1mL 1wt% trichloroauric acid aqueous solution, mix well, stir with magnetic force and boil;

[0053] (2) Quickly add 1.8mL 1wt% trisodium citrate solution, continue heating and boiling;

[0054] (3) When the color of the colloidal gold solution changes from light yellow to blue-black, and finally to wine red, continue heating and boiling for 10 minutes;

[0055] (4) When the temperature of the prepared colloidal gold solution drops to room temperature, dilute the prepared colloidal gold solution to 100mL.

[0056] (5) Finally, put the prepared colloidal gold solution in a clean glass bottle and store it at 4°C for later use.

preparation example 2

[0058] Preparation of colloidal gold-labeled SARS-CoV-2 RBD antigen

[0059] The preparation method of the colloidal gold-labeled SARS-CoV-2 RBD antigen in the application comprises the following steps:

[0060] (1) Take 2mL of the colloidal gold solution prepared in Preparation Example 1, and use 0.1M K 2 CO 3 Adjust the pH to 8.0;

[0061] (2) Add 200 μL of SARS-CoV-2 S-RBD antigen with a concentration of 0.1ug / μL dropwise, and react with magnetic stirring for 2 hours at room temperature;

[0062] (3) Add 10wt% BSA solution dropwise to make the final concentration 0.5wt%, and react with magnetic stirring for 30min at room temperature:

[0063] (4) Add 10wt% PEG20000 solution dropwise to make the final concentration 0.2wt%, and stir magnetically for 30min at room temperature;

[0064] (5) Centrifuge at 12000rpm for 30min, discard the supernatant, and leave the colloidal gold precipitate;

[0065] (6) Add 1wt% BSA solution to the precipitate to resuspend the colloidal gol...

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Abstract

The invention relates to the technical field of biomedical detection, in particular to an SARS-CoV-2 neutralizing antibody test strip, and a preparation method thereof and a kit. The test strip comprises a bottom plate as well as a sample pad, a combination pad, a detection pad and a water absorption pad which are arranged on the bottom plate and are connected in sequence; and the sample pad is coated with SARS-CoV-2 ACE2, the combination pad is coated with a colloidal gold labeled SARS-CoV-2 S-RBD antigen, the detection pad is provided with a detection line and a control line, and the detection line is coated with the SARS-CoV-2 S-RBD antigen. The test strip does not need to use related instruments, is suitable for field detection of different scenes, is short in detection time and high in detection sensitivity, and further realizes field high-accuracy rapid detection.

Description

technical field [0001] The application relates to the technical field of biomedical detection, more specifically, it relates to a SARS-CoV-2 neutralizing antibody test strip, a preparation method and a kit thereof. Background technique [0002] SARS-CoV-2 is a new type of coronavirus of the genus β, which is round or oval in shape, with a diameter of 60-140nm and a crown-like shape under an electron microscope, which can infect mammals and humans. When SARS-CoV-2 invades the body, the receptor binding domain (RBD) of the coronavirus S protein can bind the virus to angiotensin-converting enzyme 2 (ACE2) on the surface of the host cell. When the virus stimulates the B lymphocytes in the body to produce protective neutralizing antibodies, the neutralizing antibodies can bind to the antigens on the surface of pathogenic microorganisms, thereby preventing the pathogenic microorganisms from adhering to the target cell receptors and preventing them from invading cells, thereby exer...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/569G01N33/558G01N33/58G01N33/531
CPCG01N33/531G01N33/558G01N33/56983G01N33/587G01N33/6854G01N2333/165G01N2469/20
Inventor 肖潇刘国瑞黄磊童坤
Owner 南京申基医药科技有限公司
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