High-sensitivity novel coronavirus neutralizing antibody detection kit and detection method

A coronavirus and antibody detection technology, applied in the field of immunofluorescence detection, to achieve the effect of expanding the detection range, convenient operation and good linear gradient

Pending Publication Date: 2021-06-22
天津中新科炬生物制药股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In view of this, the object of the present invention is to invent a highly sensitive novel coronavirus neutralizing antibody detection kit (microfluidic method), and to invent a highly sensitive novel coronavirus neutralizing antibody indirect competitive detection method to overcome the low sensitivity of the existing new crown neutralizing antibody, accurately detect the neutralizing antibody content in the subject, and accurately evaluate the effectiveness of the new crown vaccine

Method used

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  • High-sensitivity novel coronavirus neutralizing antibody detection kit and detection method
  • High-sensitivity novel coronavirus neutralizing antibody detection kit and detection method
  • High-sensitivity novel coronavirus neutralizing antibody detection kit and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1: Preparation of a microfluidic chip for detecting novel coronavirus neutralizing antibodies

[0043] 1 Materials and Instruments

[0044] 1.1 The new crown spike protein S-RBD, ACE2 receptor protein, ACE2-Ab and S-RBD-Ab were purchased from Shenzhen Feipeng Biological Co., Ltd.;

[0045] 1.2 Avidin and EZ-link biotin were purchased from Sigma;

[0046] 1.3 Fluorescence immunoassay analyzer (F10pro), Tianjin Zhongxin Keju Biopharmaceutical Co., Ltd.

[0047] 2. Composition of chip card

[0048] The microfluidic chip for detecting neutralizing antibodies against novel coronavirus of the present invention includes: a bottom sheet and a cover sheet fixedly connected, and the substrate is sequentially provided with a marking area, a detection area and a waste liquid tank along the flow direction of the sample. The marking region of the present invention immobilizes and dries the S-RBD recombinant protein marked with fluorescent microspheres.

[0049] The T site...

Embodiment 2

[0075] Example 2: Preparation of a microfluidic chip for detecting novel coronavirus neutralizing antibodies

[0076] 1 Materials and Instruments

[0077] 1.1 The new crown spike protein S-RBD, N protein, AXL receptor protein, AXL-Ab and S-RBD-Ab were purchased from Shenzhen Feipeng Biological Co., Ltd.;

[0078] 1.2 Avidin and EZ-link biotin were purchased from Sigma;

[0079] 1.3 Fluorescence immunoassay analyzer (F10pro), Tianjin Zhongxin Keju Biopharmaceutical Co., Ltd.

[0080] 2. Composition of chip card

[0081] The microfluidic chip for detecting neutralizing antibodies against novel coronavirus of the present invention includes: a bottom sheet and a cover sheet fixedly connected, and the substrate is sequentially provided with a marking area, a detection area and a waste liquid tank along the flow direction of the sample. The marking region of the present invention is fixed and dried S-RBD and N protein marked with fluorescent microspheres. The T site of the detec...

Embodiment 3

[0107] Example 3: Preparation of a microfluidic chip for detecting novel coronavirus neutralizing antibodies

[0108] 1 Materials and Instruments

[0109] 1.1 New crown spike protein S-RBD, N protein, ACE2 receptor protein and S-RBD-Ab were purchased from Shenzhen Feipeng Biological Co., Ltd.;

[0110] 1.2 Avidin and EZ-link biotin were purchased from Sigma.

[0111] 1.3 Fluorescence immunoassay analyzer (F10pro), Tianjin Zhongxin Keju Biopharmaceutical Co., Ltd.

[0112] 2. Composition of chip card

[0113] The microfluidic chip for detecting neutralizing antibodies against novel coronavirus of the present invention includes: a bottom sheet and a cover sheet fixedly connected, and the substrate is sequentially provided with a marking area, a detection area and a waste liquid tank along the flow direction of the sample. The labeling region of the present invention is a mixture of S-RBD and N protein that are fixed and dried and labeled with fluorescent microspheres.

[011...

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Abstract

The invention provides a high-sensitivity novel coronavirus neutralizing antibody detection kit and a detection method. The high-sensitivity novel coronavirus neutralizing antibody detection kit comprises a carrier; the carrier is provided with a labeling region, a T site and a C site; the labeling region is fixedly provided with a dried signal particle labeled SARS-CoV-2 recombinant antigen; the T site is coated with a competitive substance; the competitive substance is receptor protein which specifically reacts with the signal particle labeled SARS-CoV-2 recombinant antigen; and the C site is coated with an SARS-CoV-2 recombinant antibody. According to the high-sensitivity novel coronavirus neutralizing antibody detection kit and the detection method of the present invention, the huddling effect of neutralizing antibodies can be reduced, the steric effect of the competitive substance can be increased, the combination of the neutralizing antibodies and the SARS-CoV-2 recombinant antigen can be easily achieved, the detection sensitivity and the detection accuracy can be improved, and various samples can be determined. Thedetection kit and the detection method method are suitable for different occasions, convenient to operate and bring convenient user experience.

Description

technical field [0001] The invention belongs to the technical field of immunofluorescence detection, and in particular relates to a highly sensitive novel coronavirus neutralizing antibody detection kit and detection method. Background technique [0002] Angiotensin Converting Enzyme 2 (ACE2) is a type I transmembrane protein, which is an important target for SARS-CoV-2 to infect the human body and enter cells. SARS-CoV-2 binds to ACE2 of human epithelial cells through the S-protein on the surface, and then uses intracellular substances to replicate and reproduce after invading the cells. The virus binds to angiotensin-converting enzyme 2 (ACE2) on the surface of host cells. When SARS-CoV-2 invades the body, it will stimulate the body to produce protective neutralizing antibodies, which specifically prevent pathogenic microorganisms from entering cells and prevent infection. Neutralizing antibodies are a soluble protein secreted by adaptive immune response cells, which hav...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/58G01N33/543
CPCG01N33/56983G01N33/582G01N33/543G01N2469/20G01N2333/165Y02A50/30
Inventor 李慧周洪锐魏华英李昀地刘钟泉赵娜张粲王俊水肖福磊李轩秦月贾凌云贺敬文刘陈针玉钱龙李娜孟佳李洲
Owner 天津中新科炬生物制药股份有限公司
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