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Method for preparing antigen tablet for detecting varicella-zoster virus antibody

An antibody detection, chickenpox virus technology, applied in the direction of virus/phage, biochemical equipment and methods, biological testing, etc., to achieve high qualification rate, solve the problem of complex production process, simplify the effect of preparation steps

Active Publication Date: 2015-02-25
CHANGCHUN KEYGEN BIOLOGICAL PROD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention provides a method for preparing antigen sheets for varicella virus antibody detection, which solves the problem of low quality of antigen sheets and complicated preparation process in detecting varicella antibodies by FAMA method problems, while prolonging the storage time of antigen sheets provides conditions for batch preparation of antigen sheets

Method used

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  • Method for preparing antigen tablet for detecting varicella-zoster virus antibody
  • Method for preparing antigen tablet for detecting varicella-zoster virus antibody
  • Method for preparing antigen tablet for detecting varicella-zoster virus antibody

Examples

Experimental program
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Effect test

Embodiment 1

[0024] Select Vero cells that grow to a monolayer with a density of about 2.5—3.5×10 5 / ml; varicella-zoster virus strain VZV 84-7 virus titer is 4.9lgPFU / ml; Cells are infected with a multiplicity of infection of 0.01; virus infection solution is MEM containing 1% glutamine, 2% newborn bovine serum, with 5.6 %NaHCO 3 Adjust the pH value to 7.5±0.1; place the infected cells in a constant temperature incubator at 35°C and culture until the peak of virus replication; digest with 0.25% trypsin, and count the cells; add 1% glutamine and 5% newborn bovine serum to a pH value of 7.4 ±0.1 MEM culture solution, adjust the cell concentration to 1.2~1.8×10 5 / ml to prepare cell suspension; drop the cell suspension into 12-well glass slides, 20~25μl per well, put the slides in a humid box, and put them at 35°C and 5% CO 2 After incubating in the incubator for 30 minutes, rinse with PBS once, dry naturally, and directly fix with cold acetone at room temperature for 15 minutes, take it o...

Embodiment 2

[0026] Select Vero cells that grow to a monolayer with a density of about 2.5—3.5×10 5 / ml; varicella-zoster virus strain VZV 84-7 virus titer is 5.2lgPFU / ml; Cells are infected with a multiplicity of infection of 0.01; the virus infection solution is MEM containing 1% glutamine, 2% newborn bovine serum, with 5.6 %NaHCO 3 Adjust the pH value to 7.5±0.1; place the infected cells in a constant temperature incubator at 35°C and culture until the peak of virus replication; digest with 0.25% trypsin, and count the cells; add 1% glutamine and 5% newborn bovine serum to a pH value of 7.4 ±0.1 MEM culture solution, adjust the cell concentration to 1.2~1.8×10 5 / ml to prepare cell suspension; drop the cell suspension into 12-well glass slides, 20~25μl per well, put the slides in a humid box, and put them at 35°C and 5% CO 2 After incubating in the incubator for 30 minutes, rinse with PBS once, dry naturally, and directly fix with cold acetone at room temperature for 15 minutes, take ...

Embodiment 3

[0028] Select Vero cells that grow to a monolayer with a density of about 2.5—3.5×10 5 / ml; varicella-zoster virus strain VZV 84-7 virus titer is 4.9lgPFU / ml; Cells are infected with a multiplicity of infection of 0.05; the virus infection solution is MEM containing 1% glutamine, 2% newborn bovine serum, with 5.6 %NaHCO 3 Adjust the pH value to 7.5±0.1; place the infected cells in a constant temperature incubator at 35°C and culture until the peak of virus replication; digest with 0.25% trypsin, and count the cells; add 1% glutamine and 5% newborn bovine serum to a pH value of 7.4 ±0.1 MEM culture solution, adjust the cell concentration to 1.2~1.8×10 5 / ml to prepare cell suspension; drop the cell suspension into 12-well glass slides, 20~25μl per well, put the slides in a humid box, and put them at 35°C and 5% CO 2 After incubating in the incubator for 30 minutes, rinse with PBS once, dry naturally, and directly fix with cold acetone at room temperature for 15 minutes, take ...

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Abstract

The invention provides a method for preparing an antigen tablet for detecting the varicella-zoster virus antibody. According to the method, a varicella-zoster virus VZV84-7 strain is used as an infected virus strain to prepare the antigen tablet; compared with an Oka strain infected diploid cell, the virus has the characteristics that the virus CPE is in a dispersed manifestation and is widely distributed on the surface of the cell after infecting a sensitive cell; a Vero cell is selected as a cell matrix, and can be dispersed more sufficiently than the diploid cell, and the cellular morphology is more completed; after the infected virus is reproduced, the CPE widely occurs to the cell, so that observation is facilitated and results can be determined when the virus infected cell is used for preparing the antigen tablet. By adopting the method, the problems of low antigen tablet quality and complicated tablet producing process in varicella-zoster antibody detection by an FAMA method can be solved, the preservation time of the antigen tablet can be prolonged, and conditions are provided to massive preparation of the antigen tablet. Compared with a traditional method, the method has the advantages of simplicity and practicality, the antigen tablet can be frozen at -20-70 DEG C and stored for over one year without remarkable quality change, and the result of tablet microscopic examination is not influenced.

Description

Technical field: [0001] The invention relates to a method for preparing an antigen sheet for varicella virus antibody detection. The method comprises the steps of infecting cells with varicella zoster virus to pathological changes, digesting and making a cell suspension, and then preparing the antigen sheet, which belongs to the field of biotechnology. Background technique: [0002] Currently, the commonly used detection methods for varicella-zoster virus antibody include FAMA method, mammary ball assay and ELISA method. The ELISA method is convenient and fast, and is suitable for large-scale sample detection, but in practical applications, the false negative rate is too high, and it cannot well reflect the protective defects of antibodies. The milk ball assay is relatively complicated to operate and highly subjective, so it is suitable for the detection of a small amount of samples. The FAMA method is currently internationally recognized as the gold standard for the dete...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00G01N33/68
Inventor 沈红杰李海燕郑晓丽李生军赵海波闫磊朱晓文双慧张宇收到
Owner CHANGCHUN KEYGEN BIOLOGICAL PROD
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