Construction method for suckling mouse model infected by hepatitis B virus and application of method

A technology of hepatitis B virus and its construction method, which is applied in the field of construction of suckling mouse models, can solve the problems of difficult cultivation, difficult promotion, and inability to reproduce the infection process well, and achieves the effect of low cost and no rise in body temperature

Inactive Publication Date: 2019-09-06
GENERAL HOSPITAL OF SOUTHERN THEATRE COMMAND OF PLA
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  • Application Information

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Problems solved by technology

HBV transgenic mice are widely used in drug research and gene therapy to interfere with the replication of HBV, and can even be used to study the clearance of ccc DNA. However, the cost is high, the reproducibility is average, and the research on the life cycle of HBV is not yet clear; duck hepatitis B The model is mainly aimed at avian hepatotropic virus, which is quite different from human HBV in structure, and cannot reproduce the infection process of human HBV well; tree shrews are small mammals...

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  • Construction method for suckling mouse model infected by hepatitis B virus and application of method
  • Construction method for suckling mouse model infected by hepatitis B virus and application of method

Examples

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Embodiment 1

[0033] One, a kind of construction method of the suckling mouse model of hepatitis B virus infection

[0034] Such as figure 1 As shown, it specifically includes the following steps:

[0035] 1. Extract the serum of patients with high expression of HBV infection (HBsAg, HBeAg positive HBV DNA concentration is greater than 10 5 copy / mL), placed at room temperature for 2 hours, centrifuged to obtain serum, and fluorescent quantitative PCR was used to measure serum HBV DNA concentration, and stored in a -80°C refrigerator for later use.

[0036] Five tubes of serum from HBV-infected patients were thawed and mixed, and the concentration of HBV DNA measured by real-time fluorescent quantitative PCR was 3.05×10 6 copies / mL.

[0037] 2. Three days before inoculation, blood was collected from the inner canthus vein of the mice, and HBs Ag and HBe Ag were detected by ELISA to exclude spontaneous HBV infection.

[0038] 3. 20 suckling mice were randomly divided into 4 groups, the HB...

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Abstract

The invention discloses a construction method for a suckling mouse model infected by hepatitis B virus and application of the construction method. The construction method comprises the steps of extracting serum of a patient with high expression infection, eliminating spontaneous infection of HBV in an experimental mouse, injecting the serum of the patient into the mouse by a high-pressure hydrodynamic tail vein method, respectively taking blood through an intraocular canthus vein of the mouse at 7, 15 and 30 days after injection to detect the concentration of HBs Ag, HBe Ag and HBV DNA, and determining that modeling is successful when the result is positive. The method provided by the invention can be used for successfully infecting suckling mice, it detects that the successfully modeled suckling mouse can permanently have the expression of HBV within one month, meanwhile, body temperature rising and body weight changing of the suckling mouse cannot be caused, the cost is low, ethicalproblems do not exist, the suckling mouse model can be used as an animal model for researching chronic HBV infection, and the requirements of current scientific research are met.

Description

technical field [0001] The invention relates to the technical field of animal model preparation, in particular to a construction method and application of a suckling mouse model infected with hepatitis B virus. Background technique [0002] Hepatitis B is a disease that endangers the health of all human beings. It can be transmitted through blood, iatrogenic transmission, mother-to-child transmission and sexual transmission, and can be followed by liver fibrosis, cirrhosis, liver failure, and even hepatocellular carcinoma. It is estimated that about 2 billion people in the world have been infected with HBV, of which about 400 million people are chronic hepatitis B infections. Interferon α and nucleoside analogues are currently the main antiviral drugs, which can inhibit virus replication to a certain extent, reduce inflammation and necrosis of liver cells, but cannot completely remove covalently closed circular DNA (covalent-ly closed circular DNA, ccc DNA), so chronic hepa...

Claims

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Application Information

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IPC IPC(8): A01K67/02A61K49/00
CPCA01K67/02A01K2207/00A01K2227/105A01K2267/0337A61K49/0008
Inventor 任广立
Owner GENERAL HOSPITAL OF SOUTHERN THEATRE COMMAND OF PLA
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