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Detection kit for antigens of novel coronavirus (SARS-CoV-2)

A sars-cov-2, antigen technology, applied in the direction of viruses, viral peptides, viruses/phages, etc., can solve the problems of time-consuming, cumbersome and time-consuming, difficult to guarantee specificity and sensitivity, and achieve accurate detection results and improve detection. Output rate, flexible operation effect

Inactive Publication Date: 2020-06-19
北京新创生物工程有限公司
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0002] Since the outbreak of the 2019 new coronavirus (SARS-CoV-2), both confirmed and suspected cases have grown rapidly within a certain period of time. The specific real-time fluorescent RT-PCR test results of the two targets of the new coronavirus (ORF1ab, N) in each sample are all positive to confirm that the sample is positive. , in addition, the sample preparation process is cumbersome and time-consuming, and the sample is easily contaminated during the process, which leads to great limitations of this method
In the existing technology, most of the products that claim to detect SARS-CoV-2 antigens are detected with antibodies from previous research on other coronaviruses. For example, the homology between the new crown and SARS is very high, and SARS antibodies can be used as a temporary emergency. , but the specificity and sensitivity are difficult to guarantee, which reduces the accuracy of infection detection in my country

Method used

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  • Detection kit for antigens of novel coronavirus (SARS-CoV-2)
  • Detection kit for antigens of novel coronavirus (SARS-CoV-2)
  • Detection kit for antigens of novel coronavirus (SARS-CoV-2)

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preparation example Construction

[0060] The present invention also protects the preparation method of the above-mentioned antibody, which uses a protein composed of any one of the amino acid sequences shown in SEQ ID NO.2-SEQ ID NO.11 as an antigen, prepares an immunogen with an adjuvant, and obtains an antibody after immunizing animals. Further, for example, a protein composed of any one of the amino acid sequences shown in SEQ ID NO.2-SEQ ID NO.11 is used as an antigen, and an adjuvant is used to prepare an immunogen, and after the animal is immunized, the titer of the antibody meets the 10-fold dilution activity After the level above L6, splenocytes were immunized again to perform cell fusion with myeloma cells, positive clones were screened, amplified and purified to obtain antibodies. The inventor determined the preparation scheme provided by the present invention by exploring the conditions of animal immunization, cell fusion, screening of specific hybridoma cells, and mass production of monoclonal antib...

Embodiment 1

[0075] The preparation process of embodiment 1 monoclonal antibody

[0076] animal immunity

[0077] ① Antigen preparation.

[0078] Prepare the immunization antigen of monoclonal antibody, the screening antigen of present embodiment is as follows 10 kinds: N1 antigen (SEQ ID NO.2), N2 antigen (SEQ ID NO.4), N3 antigen (SEQ ID NO.3), N4 antigen ( SEQ ID NO.5), S1 antigen (SEQ ID NO.6), S2 antigen (SEQ ID NO.7), S3 antigen (SEQ ID NO.8), M1 antigen (SEQ ID NO.9), M2 antigen (SEQ ID NO.9) ID NO.10), E antigen (SEQ ID NO.11).

[0079] ② Selection of immune animals.

[0080] Mice can be selected as immunized animals according to the myeloma cells used. Because the mouse myeloma cell lines used in hybridoma technology are all derived from BALB / c mice. Mice should be 8-12 weeks old for the first immunization, and female mice are easier to operate.

[0081] ③ Determination of the immunization program.

[0082] Choose the appropriate immunization scheme according to the specifi...

Embodiment 2

[0130] The preparation of embodiment 2 polyclonal antibody

[0131] immunity

[0132] Antigens were selected from N protein (SEQ ID NO.1), S protein (SEQ ID NO.12), M protein (SEQ ID NO.13) and E protein (SEQ ID NO.14), and polyclonal antibodies were prepared respectively.

[0133] For the first immunization, the antigen was mixed with Freund's complete adjuvant in a volume ratio of 1:1, stirred by a vortex mixer to prepare a water-in-oil adjuvant, and each guinea pig was subcutaneously immunized with multiple injections of 2ml. 0.5 mg antigen per guinea pig.

[0134] For the second immunization, 3 weeks after the first immunization, the antigen was mixed with Freund’s incomplete adjuvant in a volume ratio of 1:1, stirred by a vortex mixer to prepare a water-in-oil adjuvant, and each guinea pig was subcutaneously immunized at multiple points Inject 2ml, 0.25mg antigen per guinea pig.

[0135] For the third immunization, 3 weeks after the second immunization, take the antige...

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Abstract

The invention relates to the technical field of biology, and specifically provides a detection kit for a detection kit for antigens of a novel coronavirus (SARS-CoV-2). According to the detection kitprovided by the invention, the antigens of the SARS-CoV-2 are detected by virtue of a colloidal gold double antibody sandwich method, and two monoclonal antibodies and colloidal gold are mixed for labeling, so that a detection result of the antigens of the SARS-CoV-2 is accurate and high in sensitivity, and meanwhile, the detection rate can be remarkably increased. According to the detection kit,a blank in immunological detection of the SARS-CoV-2 is filled up, and field detection can be realized without a detection instrument, so that the time and the labor are saved, and operation is flexible.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a novel coronavirus (SARS-CoV-2) antigen detection kit. Background technique [0002] Since the outbreak of the 2019 new coronavirus (SARS-CoV-2), both confirmed and suspected cases have increased rapidly within a certain period of time. The specific real-time fluorescent RT-PCR test results of the two targets of the new coronavirus (ORF1ab, N) in each sample are all positive to confirm that the sample is positive. , In addition, the sample preparation process is cumbersome and time-consuming, and the sample is easily contaminated during the process, which leads to great limitations of this method. In the existing technology, most of the products that claim to detect SARS-CoV-2 antigens are detected with antibodies from previous research on other coronaviruses. For example, the homology between the new crown and SARS is very high, and SARS antibodies can be used as a temporary emerg...

Claims

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Application Information

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IPC IPC(8): C07K14/165C07K16/10C12N5/20G01N33/543G01N33/558G01N33/569
CPCC07K14/005C07K16/10C12N2770/20022G01N33/54313G01N33/558G01N33/56983
Inventor 景滢滢刘兴旺牛犇姜化冰柴茜
Owner 北京新创生物工程有限公司
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