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Method for predicting titer of novel coronavirus neutralizing antibody, and kit thereof

A coronavirus and antibody titer technology, which is applied in the field of neutralizing antibody titer prediction, can solve the problems of inability to meet the requirements of rapid, high-throughput, accurate detection, inability to simulate and replace live virus infecting cells, and complicated operations, and avoid False positive samples are falsely detected, the specificity can be quantitatively detected, and the operation is simple.

Active Publication Date: 2021-04-06
杭州凯曼健康科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

PRNT detection uses live viruses and needs to be completed in a BSL-3 laboratory, and the PRNT detection operation is complicated, the cycle is long, and the error is large
Although pseudovirus neutralization detection uses a highly safe pseudovirus, it cannot simulate and replace the process of live virus infecting cells, and it also has complex operations, long cycles, and large errors, which cannot meet the needs of rapid, high-throughput, and accurate detection. needs

Method used

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  • Method for predicting titer of novel coronavirus neutralizing antibody, and kit thereof
  • Method for predicting titer of novel coronavirus neutralizing antibody, and kit thereof
  • Method for predicting titer of novel coronavirus neutralizing antibody, and kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] Preparation of novel coronavirus recombinant antigen N protein, S1-RBD protein and S2-ECD protein:

[0071] (1) Using gene cloning technology, PCR amplifies the N gene, S2-ECD gene, and S1-RBD protein encoding the novel coronavirus antigen;

[0072] (2), N gene is transferred into Escherichia coli for expression, S2-ECD gene is transferred into 293 cells for expression, S1-RBD protein is transferred into 293 cells for expression, and new coronavirus recombinant antigen N protein and S2-ECD protein are obtained and S1-RBD protein.

Embodiment 2

[0074] Preparation of chromatographic reaction membrane (two detection lines of IgA and IgG):

[0075] Paste the NC membrane with a width of 25cm on the middle area of ​​the PC bottom card, and then dilute the anti-human IgA, anti-human IgG and goat anti-rabbit IgG antibodies with PBS diluent to 0.2-1mg / ml respectively and draw them with a gold sprayer. In the corresponding detection line area and quality control line area of ​​the NC membrane. Dry in an oven at 37°C for 12-24 hours, and seal the bag for later use.

Embodiment 3

[0077] Preparation of chromatographic reaction membrane (two detection lines of IgM and IgG):

[0078] Paste the NC membrane with a width of 25cm on the middle area of ​​the PC bottom card, and then dilute the anti-human IgM, anti-human IgG and goat anti-rabbit IgG antibodies with PBS diluent to 0.2-1mg / ml respectively and draw them with a gold sprayer In the corresponding detection line area and quality control line area of ​​the NC membrane. Dry in an oven at 37°C for 12-24 hours, and seal the bag for later use.

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Abstract

The invention provides a method for predicting titer of a novel coronavirus neutralizing antibody, and a kit thereof, and relates to the technical field of titer prediction of neutralizing antibodies. The method for predicting titer of the novel coronavirus neutralizing antibody comprises the following steps: firstly, providing a biological sample to be detected; then detecting the expression level of the novel coronavirus specific protein in a serum sample after being combined with an affinity antibody; and associating the detection result with the neutralizing antibody titer to predict the neutralizing antibody titer of the novel coronavirus in the biological sample to be detected. According to the method, the neutralizing antibody result of the novel coronavirus can be well predicted only through serum detection, and compared with existing plaque reduction neutralizing tests and pseudovirus neutralizing titer detection, and the method has the advantages of being easy to operate, high in detection speed, short in detection period and safe in detection environment, and can be used as an evaluation index for secondary recurrence after novel coronavirus vaccination or infection.

Description

technical field [0001] The invention relates to the technical field of neutralizing antibody titer prediction, in particular to a method for predicting a novel coronavirus neutralizing antibody titer and a kit thereof. Background technique [0002] After vaccination, the human body can produce protective antibodies, namely neutralizing antibodies, through the immune response. The titer of the neutralizing antibody titer can be used to judge the protective effect of the vaccine. The detection of neutralizing antibody titer in COVID-19 patients after recovery can be used to determine whether there is a risk of secondary infection. [0003] Currently established coronavirus neutralizing antibody titer detection methods include plaque reduction neutralization test (Plaque Reduction Neutralization Test, PRNT) and pseudovirus neutralization titer detection (Pseudovirus Based Neutralization Assay, PBNA). PRNT detection uses live viruses and needs to be completed in a BSL-3 labora...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/58G01N33/558G01N33/543G01N33/68
CPCG01N33/56983G01N33/588G01N33/558G01N33/543G01N33/6854G01N2333/165G01N2469/20
Inventor 姚航平陈喆陈杭曹少伟陈秋强沈明程吴云
Owner 杭州凯曼健康科技有限公司