动物造模,腺病毒 z-bio 2022-06-30 493

　　Objective: To understand the natural infection of mouse adenovirus (Mad) in our province and explore the distribution of virus in various organs and tissues of artificially infected Mad mice and the changes of serum antibodies.

　　Method: The ELISA method was used to detect the virus antibody in the serum of mice raised in ordinary environments in 2007 and 2015, and the serum of SPF mice from 12 supervisory units and 32 commissioned units in Guangdong Province from 2013 to 2015. Infect 36 3-week-old BALB/c mice by intraperitoneal injection, 0.2 mL/mouse, at a concentration of 4.5×106 copies/μL. Observe the clinical manifestations of the animals every day, and on the 0th day before and 3rd after the challenge , 7, 10, 15, 18, 21, 30, 37, 44, 51, 60 days of necropsy mice (3 each), take tissue samples (heart, liver, spleen, lung, kidney, brain, stomach, cecum Contents) and serum, real-time fluorescent quantitative PCR (QPCR) method is used to detect tissue virus nucleic acid, and ELISA method is used to detect serum antibody level.

　　Result: The positive rate of antibodies in common reared mice is 24.44%-84.15%, among which the serotype of Mad-2 K87 strain is the main one. The positive rate of SPF mouse Mad serum antibody was 0%. All challenged mice showed insidious infection and no clinical manifestations. On the 3rd and 7th day after the challenge, the positive rate of viral nucleic acid detection in each tissue of the mouse was 100% (3/3), and the 100% positive rate of the spleen maintained the longest time (60 days). Except for the liver, the virus content in each tissue of the mouse reached a peak 7 days after the challenge. The virus content in the spleen was the highest (5.5×105 copies/μL), followed by the heart (3.4×105 copies/μL), and the contents of the cecum ( 2.6×105 copies/μL) and stomach (2.6×105 copies/μL), brain is 0.8×105 copies/μL. Serum antibodies can be detected 15 days after the challenge, reaching a peak at 37 days, and maintaining a high level until 60 days thereafter.

　　Conclusion: The Mad infection rate of SPF mice is low, and the Mad infection rate of mice raised in a normal environment is high. The artificially infected mice have the highest virus content in the spleen, and the positive rate lasts the longest, indicating that the virus can replicate in the spleen for a long time. The 7th day after infection is the best time point for tissue viral nucleic acid detection. Serum antibodies can be used as monitoring indicators within 15 to 60 days after infection.