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Construction method of chronic hepatitis E virus mouse model

A hepatitis E virus and mouse model technology, applied in the field of biomedicine, can solve the problems of lack of animal models for HEV chronic infection and limited research on pathogenic mechanism, and achieve the effect of convenient breeding, low price and simple operation

Active Publication Date: 2019-05-21
KUNMING UNIV OF SCI & TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Due to the lack of suitable animal models for the study of HEV chronic infection, the study of its pathogenic mechanism is limited

Method used

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  • Construction method of chronic hepatitis E virus mouse model
  • Construction method of chronic hepatitis E virus mouse model
  • Construction method of chronic hepatitis E virus mouse model

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Experimental program
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Effect test

Embodiment 1

[0029] Embodiment 1: The construction method of the chronic hepatitis E virus mouse model is as follows:

[0030] 1. Collect feces from rhesus monkeys chronically infected for 93 weeks, prepare a 10% mass-volume ratio of PBS (pH7.4) feces suspension, shake vigorously to emulsify the feces, centrifuge at 12,000 g for 10 minutes at 4°C, and collect clear, 0.45 μm and 0.22 μm filter membrane to filter and sterilize, add double antibody solution (400U / mL penicillin and 1000U / mL streptomycin) of 5% of the filtrate volume to the filtrate, treat at 4 ℃ for 2 h, store in liquid nitrogen for later use, The virus copy number was determined to be 2.4×10 by Real-time qPCR 4 Copy number / mL;

[0031] 2. Inoculate the feces suspension treated with the above double antibody solution into SPF grade BALB / c mice through the tail vein;

[0032] 3. Collect feces and serum once a week for the detection of HEV RNA and the calculation of virus copies;

[0033] Fecal HEV RNA was extracted as follow...

Embodiment 2

[0038] Embodiment 2: The construction method of the chronic hepatitis E virus mouse model is as follows:

[0039] (1) Prepare a feces suspension in PBS (pH7.4) with a mass volume concentration of 15% from the feces of rhesus monkeys chronically infected with HEV, oscillate to emulsify the feces, centrifuge at 6°C and 10,000g for 10 minutes, collect the supernatant, and use 0.45 μm filter membrane and 0.22 μm filter membrane filter to sterilize, add double antibody solution (400U / mL penicillin and 1000U / mL streptomycin) in the filtrate to the filtrate, treat at 4°C for 2 h, store in liquid nitrogen for later use , the virus copy number was determined to be 2.1×10 by Real-time qPCR 5 Copy number / mL;

[0040] (2) Inject the feces suspension treated with the above double antibody solution into SPF grade BALB / c mice through the tail vein;

[0041] (3) Collect feces and serum every week for HEV RNA detection and virus copy detection;

[0042] The results showed that HEV RNA was c...

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Abstract

The invention discloses a construction method of a chronic hepatitis E virus mouse model. In the method, HEV strains are derived from feces of chronically infected rhesus monkeys, and the rhesus monkeys continuously expel toxin for 93 weeks; after an HEV virus suspension is inoculated into BALB / c mice through caudal veins, HEV RNA can be continuously detected in feces and serum of the mice, and the HEV RNA has high virus copying property, and the continuous positive time can reach 36 weeks. The method for establishing the model is simple, can be used for researching the persistent infection mechanism of HEV, and can provide an important animal model for screening and evaluation of anti-HEV drugs.

Description

technical field [0001] The invention belongs to the field of biomedicine, in particular to a method for establishing a hepatitis E virus (HEV) chronic mouse model. Background technique [0002] Hepatitis E Virus (HEV) is a viral hepatitis pathogen transmitted through the intestinal tract, which can infect humans and various animals across species. HEV is a single-stranded positive-sense RNA virus with a genome length of approximately 7.3kb, consisting of three open reading frames (ORF1, ORF2, and ORF3). Current studies have found that HEV mainly has 8 genotypes (HEV-1~HEV-8), and the genotype 4 HEV strain (HEV-4) is mainly prevalent in China. It has been confirmed that the transmission route of HEV is mainly the fecal-oral route, and it can also be transmitted by other routes, such as blood transmission, contact transmission, vertical transmission and organ transplantation. [0003] HEV infection is generally an acute self-limiting infection, and patients can recover on th...

Claims

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Application Information

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IPC IPC(8): A01K67/02A61K35/76
Inventor 黄芬郝先辉禹文海李云龙
Owner KUNMING UNIV OF SCI & TECH
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