动物造模,小鼠脑脊髓炎病毒 z-bio 2021-10-08 563

　　Objective: To establish a RT-PCR method for mouse encephalomyelitis virus and apply it preliminarily.

　　Methods: Design specific primers based on the genome sequence of the TMEVGDVII strain (GI: 62039) published by NCBI, establish an RT-PCR method, verify the sensitivity and specificity of the method, and check the brain cavity inoculation method 9 BALB/c mice and samples After 6 days, samples of animal brain, heart, liver, spleen, lung, kidney, appendicitis contents and serum were taken, and the established method was used. At the same time, 100 samples were tested for the contents of appendicitis in mice and the method was preliminarily applied.

　　Results: Using GDVIINA as a template, the established RT-PCR method can amplify a single band of about 371 bp. The sensitivity verification shows that the minimum detectable GDVII cDNA is 0.69 pg/μL. Explain that there is. And with lymphocytic choriomeningitis virus, Japanese encephalitis virus, murine norovirus and normal mouse brain tissue as controls for method specific verification, no target bands appeared. After the infected mice were inoculated into the brain cavity, all mice experienced varying degrees of discomfort and hind limb paralysis on the third day of inoculation. On the fifth day, 2 mice died (heart, liver, spleen, lung, kidney, brain). GDVIINA was detected in the brain tissues of 100 mice, but no other tissues were detected. The contents of the cecum of 100 mice were tested, and the results were all negative.

　　Conclusion: The T-PCR method established by the TMEVGDVII strain can effectively detect viral mouse tissue infections and can be used as a powerful supplement to the national standards for experimental animals.