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Establishment method of human rotavirus induced pathological damage and diarrhoea suckling mouse animal models

A human-derived rotavirus and rotavirus technology, applied in biochemical equipment and methods, virus/bacteriophage, microorganism measurement/inspection, etc., can solve the problem of expensive experimental animals, high feeding conditions, and low success rate of modeling. For advanced problems, achieve short modeling time, high modeling success, and stable observed effects

Inactive Publication Date: 2018-01-09
SHANDONG YIDU BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] At present, with regard to rotavirus animal models, there are rats, rabbits, gnotobiotic piglets, sterile cows, and primates as experimental animals, which exist or the success rate of modeling is not high, or the experimental animals are expensive, High feeding conditions

Method used

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  • Establishment method of human rotavirus induced pathological damage and diarrhoea suckling mouse animal models
  • Establishment method of human rotavirus induced pathological damage and diarrhoea suckling mouse animal models
  • Establishment method of human rotavirus induced pathological damage and diarrhoea suckling mouse animal models

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Experimental program
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Embodiment Construction

[0026] (1) Rotavirus culture

[0027] 1. Cell culture:

[0028] Vero cells were grown and passaged in IMDM medium containing 10% fetal bovine serum.

[0029] 2. Virus amplification:

[0030] Vero cells were inoculated with human rotavirus (WA strain) when grown to a monolayer in culture flasks. Take the WA strain of human rotavirus out of the -60°C refrigerator and dissolve at room temperature. The Vero cells grown into a single layer were rinsed twice with sterile PBS with a pH of 7.2, inoculated with the virus solution, and added with a virus maintenance solution containing trypsin, incubated at 36.5°C with 5% carbon dioxide for 2 hours, then discarded the virus solution, PBS Rinse twice, add trypsin-containing, serum-free IMDM medium. Then place it at 36.5°C and 5% carbon dioxide to culture and observe. When the virus-infected Vero cell lesion reaches +++~++++, put the culture bottle into -60°C for freezing, and then thaw at 4°C, and freeze and thaw repeatedly. Centrif...

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Abstract

The invention provides an establishment method of human rotavirus induced pathological damage and diarrhoea suckling mouse animal models. The establishment method comprises following steps: SPF grade4 to 5 days old NIH sucking mice with weight ranging from 3.5g to 5.0 are randomly selected as experimental animals; animal waste is collected, and colloidal gold method is adopted for rotavirus antigen detection; 10 of the sucking mice with negative results are selected so as to obtain a model group, each of the 10 sucking mice is fed with 0.2ml of human rotavirus centrifugation harvested fluid through the oral cavities; 10 of the sucking mice with negative results are selected so as to obtain a reference group, each of the 10 sucking mice is fed with 0.2ml of Vero cell freeze thawing centrifugation supernate through the oral cavities; 8h later, rotavirus antigen in the animal waste of the model group is detected, if obtained results are positive, establishment of the rotavirus infected sucking mouse experimental model is realized; gold labeled method is adopted to detect rotavirus antigen in the animal waste of the reference group, and the results are negative. The advantages of theestablishment method are that: simulation of rotavirus infected acute gastroenteritis characteristics of infants and children such as diarrhoea can be realized, and rapid establishment of rotavirus infected sucking mouse experimental models can be realized.

Description

technical field [0001] The invention relates to an animal model establishment method for virus strain virulence analysis, in particular to an animal model establishment method for human rotavirus research strain virulence analysis. Background technique [0002] At present, with regard to rotavirus animal models, there are rats, rabbits, gnotobiotic piglets, sterile cows, and primates as experimental animals, which exist or the success rate of modeling is not high, or the experimental animals are expensive, High feeding conditions. Therefore, it is necessary to establish an experimental animal model that is stable, easy to operate, cost-effective, and conforms to the characteristics of infantile rotavirus acute gastroenteritis. Contents of the invention [0003] The object of the present invention is to solve the problems in the above-mentioned prior art, and provide a rotavirus-infected infant rotavirus-inducing process that can simulate the symptoms of acute gastroenteri...

Claims

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Application Information

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IPC IPC(8): A61K35/76C12N7/00C12Q1/70
Inventor 樊可
Owner SHANDONG YIDU BIOTECH
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