Fluorescence-labeled varicella-zoster virus immunochromatography test paper and application thereof

A herpes zoster virus, immunochromatographic detection technology, applied in the direction of antiviral immunoglobulin, application, viral peptides, etc., can solve the problems of high detection equipment requirements, unsuitable basic laboratories, long detection cycle, etc., to achieve accurate virus detection. Antigen content, guiding vaccine development, and the effect of rapid diagnosis

Pending Publication Date: 2022-04-12
ZHENGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0004] At present, the traditional method (pharmacopoeia method) for detecting VZV virus is the plaque method, but this method has a long detection cycle, high detection requirements, and cumbersome operation, and is not suitable for basic-level laboratories; PCR amplification method can also be used to detect VZV virus Titer method, but this method has higher requirements for detection equipment

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  • Fluorescence-labeled varicella-zoster virus immunochromatography test paper and application thereof
  • Fluorescence-labeled varicella-zoster virus immunochromatography test paper and application thereof
  • Fluorescence-labeled varicella-zoster virus immunochromatography test paper and application thereof

Examples

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Embodiment 1

[0029] The specific embodiments of the present invention will be described in further detail below in conjunction with the examples, but the protection scope of the present invention is not limited to this; the instruments and equipment involved in the following examples are conventional instruments and equipment unless otherwise specified; the reagents involved Unless otherwise specified, they are all commercially available conventional reagents; the test methods involved are conventional methods unless otherwise specified. Example 1 Expression and purification of VZV gE protein

[0030] The VZV glycoprotein gE is the most abundant on the viral envelope, is the main antigen of the virus, and is also the main candidate antigen for the preparation of virus subunit vaccines and detection reagents. The expression of VZV gE protein and the preparation of monoclonal antibody are of great significance to the development of subunit vaccine and detection reagents for herpes zoster. M...

Embodiment 2

[0043] Example 2 Preparation of monoclonal antibodies

[0044] 1. Animal immunity

[0045] (1) adding Freund's complete adjuvant to the immunogen gE protein (gE protein purified in Example 1), and emulsified for the first immunization;

[0046] (2) 2 female BALB / c mice aged 4-8 weeks were immunized by subcutaneous injection at multiple points on the back, and the immunization dose was 10 μg / mice;

[0047] (3) BALB / c mice were boosted with the same method and dose after emulsification with incomplete Freund's adjuvant and immunizing antigen (gE protein purified in Example 1) every 2 weeks, and immunized 4 times in total;

[0048] (4) After 4 immunization, the tail vein blood was collected to measure the specific antibody titer against gE protein, and the mice with higher titer were selected, and 3 to 4 days before cell fusion, the method of tail vein injection was carried out, with no adjuvant. BALB / c mice were super-immunized with a dose of immunogen, and the immunization do...

Embodiment 3

[0067] Example 3 Purification and identification of antibodies

[0068] 1. Purify antibody by saturated ammonium sulfate precipitation method. The operation method is as follows:

[0069] (1) Take 5 mL of monoclonal antibody ascites (obtained in Example 2), add 5 mL of PBS buffer, and then add 2.5 mL of saturated ammonium sulfate solution dropwise to make a final concentration of 20% (wt%) ammonium sulfate solution, while adding While stirring, after fully mixing, let stand for 30min.

[0070] (2) 8000r / min, centrifuge for 20min, discard the precipitate to remove fibrin.

[0071] (3) Add 12.5 mL of saturated ammonium sulfate solution to the supernatant, mix well, and let stand for 30 minutes.

[0072] (4) Centrifuge at 8000 r / min for 20 min, and discard the supernatant.

[0073] (5) Add 10 mL of PBS buffer to the precipitate to dissolve it, and then add 5 mL of saturated ammonium sulfate solution to make it into a 33% (wt%) ammonium sulfate solution, mix well, and let stand...

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Abstract

The invention provides immunochromatography detection test paper for a fluorescently-labeled varicella-zoster virus and application of the immunochromatography detection test paper. The test paper comprises a supporting bottom plate and an adsorption layer fixed on the supporting bottom plate, wherein the adsorption layer sequentially comprises a sample pad, a combination pad, a nitrocellulose membrane and a water absorption pad from a test end; the nitrocellulose membrane contains a detection line T and a quality control line print; a monoclonal antibody 7E12 marked by quantum dots is marked on the combination pad; a monoclonal antibody 5B10 is marked by the detection line T; sPA is marked by the quality control line. Wherein the monoclonal antibody 7E12 and the monoclonal antibody 5B10 are prepared by using a gE protein as an antigen. The glycoprotein gE monoclonal antibody can specifically recognize and resist the varicella-zoster virus glycoprotein gE protein and the varicella-zoster virus, and can be used for detecting the varicella-zoster virus glycoprotein gE and the varicella-zoster virus.

Description

technical field [0001] The invention relates to a fluorescently labeled varicella-zoster virus immunochromatographic detection test paper and application thereof, belonging to the technical field of biological detection. Background technique [0002] Varicella-zoster virus (varicella-zoster virus, VZV) refers to the varicella-zoster virus (varicella-zoster virus, VZV) that causes chickenpox in children for the first time, and the virus is latent in the body after recovery. herpes zoster virus. Chickenpox is a highly contagious common disease in children. It usually occurs between 2 and 6 years old. The source of infection is mainly patients. The contents and respiratory secretions of chickenpox in the acute stage of patients contain the virus. [0003] In recent years, studies at home and abroad have found that the number of clinical complications of chickenpox has increased significantly. Chickenpox has increasingly become a disease with the highest incidence among childre...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/569G01N33/558G01N33/52C07K14/04C07K16/08C12N15/867C12N15/38
Inventor 王爱萍牛艳周景明丁培阳朱习芳张盈祁艳华李泽慧
Owner ZHENGZHOU UNIV
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