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Rotavirus challenge animal model and establishment method thereof

A rotavirus and animal model technology, applied in the biological field, can solve the problems of low incidence of rotavirus, inability to complete the immunization process, and hinder the construction of rotavirus challenge model of older suckling mice, so as to improve the stability of the model Sexuality and the effect of increasing the incidence of rotavirus

Active Publication Date: 2022-02-01
BEIJING CELL FUSION BIOTECHNOLOGY CO LTD +1
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since the vaccine immunization has gone through at least one, two or more immunization procedures, and 7-day-old suckling mice are too young to complete the immunization process, the animal model of 7-day-old suckling mice cannot be used to evaluate the efficacy of rotavirus vaccines. The immune effect, therefore, a challenge model of older suckling mice is needed to evaluate the immune effect of the vaccine
[0004] However, the age of suckling mice has a significant impact on the incidence of rotavirus, and the older the suckling mice, the lower the incidence of rotavirus, which seriously hinders the establishment of a rotavirus challenge model in older suckling mice Therefore, it is urgent to find a method to improve the incidence of rotavirus in older suckling mice and the stability of the model, so as to evaluate the effectiveness of rotavirus vaccine

Method used

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  • Rotavirus challenge animal model and establishment method thereof
  • Rotavirus challenge animal model and establishment method thereof
  • Rotavirus challenge animal model and establishment method thereof

Examples

Experimental program
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Effect test

preparation example Construction

[0063] The preparation method of rotavirus liquid in the following examples is as follows:

[0064] Preparation of monolayer MA104 cells: Resuscitate MA104 cells (purchased from China Center for Type Culture Collection) into a T25 cell bottle filled with DMEM medium (purchased from Gibco), culture at 37°C for 24 hours, discard the old culture Replace with new DMEM medium, and continue to culture at 37°C for 3 days to obtain monolayer MA104 cells; use DMEM medium to subculture the monolayer MA104 cells in T75 cell flasks at a ratio of 1:3 Cultivate, after subculturing for 3 days, the T75 cell flask is covered with a single layer of MA104 cells, which is used for rotavirus inoculation;

[0065] Activation of rotavirus: Take 1 mL of rotavirus virus liquid (rotaviruses are SA11 strain purchased from ATCC and CDC-9 strain purchased from US CDC respectively), and add a final concentration of 15 μg / mL to the virus liquid Trypsin (purchased from Gibco) and calcium chloride (purchased...

Embodiment 1

[0079] Embodiment 1: The influence of age on the incidence of Kunming mouse suckling mice

[0080] The experimental animals were Kunming mouse suckling rats (purchased from Beijing Weitong Lihua Company). There were 5 experimental groups in total. After 1 hour of starvation, intragastric administration was carried out to attack the poison, and the dose of the poison was SA11 6.75lgCCID 50 / only, using a No. 6 gavage needle. After the virus attack, the diarrhea situation was counted every day and scored according to the degree of diarrhea.

[0081] According to the collected data, draw the change curve of diarrhea rate and the change curve of diarrhea score, which can be seen in detail Figure 1~2; Among them, the formula for calculating the diarrhea rate is: diarrhea rate (%) = number of suckling mice with diarrhea / total number of suckling mice × 100%; the evaluation method for the degree of diarrhea is: observe the shape of suckling rat feces and score, divided into 4 grad...

Embodiment 2

[0083] Embodiment 2: Effects of different challenge doses on the incidence of suckling mice

[0084] The experimental animals were 7-day-old Kunming mouse suckling mice (purchased from Beijing Weitong Lihua Co., Ltd.). There were 4 groups in the experimental group, of which 3 groups of suckling mice were starved for 1 hour and then intragastrically challenged. The challenge doses were CDC -9 5.50 lgCCID 50 / only, 6.50lgCCID 50 / only, 7.25 lgCCID 50 / mouse, the remaining 1 group of suckling mice was starved for 1 h and then gavaged with the same volume of PBS buffer as a negative control (Mock). After the virus attack, the diarrhea situation was counted every day and scored according to the degree of diarrhea.

[0085] According to the collected data, the change curve of diarrhea rate and the change curve of diarrhea score can be seen in detail Figure 3~4 ; Among them, the formula for calculating the diarrhea rate is: diarrhea rate (%) = number of suckling mice with diar...

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Abstract

The invention relates to a rotavirus challenge animal model and an establishment method thereof, and belongs to the technical field of biology. The invention provides a rotavirus challenge animal model. The rotavirus challenge animal model is a suckling mouse filled with an anti-acid protective agent (such as disodium adipate, sodium bicarbonate, sodium succinate, sodium citrate, sodium malate and sodium acetate) and rotavirus. Researches find that before or when the rotavirus is used for counteracting toxic substances, the rotavirus morbidity and the model stability of the large-day-old suckling rats can be effectively improved by irrigating the large-day-old suckling rats with the anti-acid protective agent; the large-day-old suckling mouse can complete the complete immune process of the vaccine; therefore, besides the virulence evaluation of the rotavirus strain, the rotavirus challenge animal model also has an application prospect in the evaluation of the effectiveness of the rotavirus vaccine.

Description

technical field [0001] The invention relates to a rotavirus challenged animal model and a method for establishing the same, belonging to the field of biotechnology. Background technique [0002] Rotavirus (RV) is a double-stranded RNA virus belonging to the Reoviridae family. It is one of the main pathogens that cause diarrhea in infants and young children under the age of five. It infects hundreds of millions of people worldwide every year and causes dozens of deaths. Ten thousand. In my country, about 10 million infants suffer from rotavirus-infected gastroenteritis every year, accounting for 1 / 4 of the number of infants. It is the most important pathogen causing severe diarrhea in infants and young children. Rotavirus was first discovered in 1973, but there is still no specific drug, and the protective effects of existing rotavirus vaccines vary greatly in different regions and there are safety problems. Therefore, research on the pathogenic mechanism of rotavirus and T...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K67/02A61K49/00
CPCA01K67/02A61K49/0008A01K2267/03A01K2227/105
Inventor 安祺张雅春田大勇王金亮
Owner BEIJING CELL FUSION BIOTECHNOLOGY CO LTD
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