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A test strip for joint detection of IgG, IGM and neutralizing antibody, its preparation method and kit

A combined detection and test strip technology, applied in the field of biomedical detection, can solve the problems of long detection time, cumbersome operation, and low detection efficiency of detection personnel, and achieve the effect of reducing detection work and high-accuracy and fast detection

Active Publication Date: 2021-10-08
南京申基医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If the inspector needs to determine whether the production of antibodies in the body is due to virus infection or vaccination, they need to use IgG / IgM test strips to select the body with IgG / IgM first, and then perform neutralizing antibody detection on this part of the body, and then The operation is cumbersome and the detection time is long, which makes the detection efficiency of the detection personnel low

Method used

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  • A test strip for joint detection of IgG, IGM and neutralizing antibody, its preparation method and kit
  • A test strip for joint detection of IgG, IGM and neutralizing antibody, its preparation method and kit
  • A test strip for joint detection of IgG, IGM and neutralizing antibody, its preparation method and kit

Examples

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preparation example Construction

[0052] Examples of Preparation of Raw Materials and / or Intermediates

[0053] The raw materials in this application can use commercially available products, and the following sources are only examples, and do not mean that they are designated raw materials.

[0054] Chicken IgY was purchased from Shanghai Yanmu Industrial Co., Ltd. (Cat. No. FK-025).

[0055] 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, pH7.4.

[0056] Anti-human IgG was purchased from Shanghai Jianglai Biotechnology Co., Ltd., the article number is AbM59504-10-PU.

[0057] Anti-human IgM was purchased from Merck anti-human anti-human IgM (μ-chain specific) goat antibody, Cat. No. 10759.

[0058] The SARS-CoV-2 N antigen was purchased from Nanjing Shenji Biotechnology Co., Ltd., the article num...

preparation example 1

[0063] Preparation of colloidal gold

[0064] The preparation method of colloidal gold in the present application adopts sodium citrate reduction method to prepare colloidal nanoparticles, comprising the following steps:

[0065] (1) Take 99mL ultrapure water and 1mL 1wt% trichloroauric acid aqueous solution, mix well, stir with magnetic force and boil;

[0066] (2) Quickly add 1.8mL 1wt% trisodium citrate solution, continue heating and boiling;

[0067] (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 8 minutes;

[0068] (4) When the temperature of the prepared colloidal gold solution drops to room temperature, dilute the prepared colloidal gold solution to 100mL;

[0069] (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

[0071] Preparation of colloidal gold-labeled SARS-COV-2 N antigen

[0072] (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;

[0073] (2) Add 200 μL of SARS-COV-2 N antigen with a concentration of 0.1 μg / μL dropwise, and react with magnetic stirring for 2 hours at room temperature;

[0074] (3) Add 10wt% BSA solution dropwise to make the final concentration 0.5wt%, and react with magnetic stirring for 30min at room temperature:

[0075] (4) Add 10wt% PEG20000 solution dropwise to make the final concentration 0.2wt%, and stir magnetically for 30min at room temperature;

[0076] (5) Centrifuge at 12000rpm for 30min, discard the supernatant, and leave the colloidal gold precipitate;

[0077] Add 1wt% BSA solution to the precipitate to resuspend the colloidal gold, and then centrifuge at 12000rpm. ) to resuspend the pellet to 1 / 10 of the original volume to prepare colloidal gold-labeled SARS-COV-2 N anti...

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Abstract

The application relates to the technical field of biomedical detection, in particular to a test strip for joint detection of IgG, IgM and neutralizing antibodies, a preparation method and a kit thereof. The test strip includes a first detection part and a second detection part, both of which include a bottom plate and a sample pad, a binding pad, a detection pad and a water-absorbing pad arranged on the bottom plate and connected in sequence; the binding pad of the first detection part N antigen and S antigen labeled with colloidal gold, the detection pad of the first detection part is provided with a first detection line and a second detection line; the sample pad of the second detection part is coated with ACE2, the Colloidal gold-labeled S-RBD antigen is coated on the binding pad of the second detection part, and a third detection line is arranged on the detection pad of the second detection part. The application can realize on-site high-accuracy rapid detection, and the detection personnel only need to use a test strip to accurately measure IgG, IgM and neutralizing antibodies in the body.

Description

technical field [0001] The application relates to the technical field of biomedical detection, more specifically, it relates to a test strip for combined detection of IgG, IgM and neutralizing antibodies, a preparation method and a kit thereof. 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. The research results show that it is highly infectious and pathogenic, and currently there is no specific clinical drug against SARS-CoV-2 infection. To this end, cutting off the source of infection from the source, suppressing the spread of the virus, and promoting the development and vaccination of vaccines are effective ways to effectively control the spread of the virus at present, and timely and effective detection of whether there is a new type of coronavirus in the body and ensuring the effe...

Claims

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Application Information

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