动物造模,减蛋综合征 z-bio 2021-11-16 456

　　Objective: Through artificial infection with egg drop syndrome virus (EDSV), observe the proliferation and dynamic changes of the virus in different strains of mice, and provide theoretical basis and data support for the construction of EDSV vectors.

　　Methods: BALB/c mice with normal immune system, nude mice with T cell immunodeficiency (Nu) and highly immunodeficient mice (NSG) were selected as the research objects, 32 per strain, female, 5-6 weeks old, intraperitoneal EDSV was artificially infected by injection, and serum was collected at 1st, 3rd, 5th, 7th, 14th, 21, 28, and 35 days after the challenge, and the indirect ELISA method was used for antibody monitoring; select 1st, 7th, 14th, 14th, 21st, 28th day after challenge. Rats, collect heart, lung, liver, spleen, kidney, small intestine, uterus, trachea, esophagus, and brain 10 types of tissues, and use fluorescence quantitative PCR relative quantitative comparison Ct method (△△CT) to detect the viral load in each tissue.

　　Results: BALB/c mice could detect antibody expression in the serum 3 days after challenge, and the antibody level reached the highest at 14 days and maintained until 35 days during the monitoring period; Nu mice could also be detected 3 days after challenge The expression level of antibody was lower than that of BALB/c mice. After 14 days of challenge, the antibody level in the serum of Nu mice decreased, and the antibody remained at a low level until 35 days. In the entire monitoring process of NSG mice, the antibody level The level has been negative. Nucleic acid relative quantitative results showed that 1 day after BALB/c mice were infected, the virus expression level in liver tissue was the highest, reaching 5.45 orders of magnitude, followed by the spleen, esophagus, uterus, small intestine, from high to low. The lung, trachea, kidney, heart, and brain tissues have the lowest levels of virus. As the infection time increases, the level of virus expression in each tissue decreases compared with 1 day after infection. By 28 days after the challenge, the level of liver and spleen virus expression is still maintained. Higher levels; Nu mice and NSG mice were infected with the highest virus expression in the spleen on the first day, which was 3.95 and 4.05 orders of magnitude, respectively, followed by the liver, which was challenged for 28 days. The organs of the two mice can still be detected Positive signal, high expression of liver and spleen viruses.

　　Conclusion: EDSV can stimulate the immune response in mice, and the expression level of antibody is low in immunodeficient mice. The virus has tissue tropism such as liver and spleen in mice. It has been developed as a vector for EDSV and used in experimental animal models. Provide reference data for further research and application.