动物造模,多瘤病毒,荧光PCR检测 z-bio 2021-12-23 385

　　Objective: To establish a fluorescent quantitative PCR detection method for polyoma virus, and to study polyoma virus infection in nude mice.

　　Methods: Compare the nucleotide sequence of mouse polyoma virus (Genbank: NC_001515) with NCBI, select storage area, design primers and probes, establish a fluorescent quantitative PCR method for polyoma virus, and verify the sensitivity of the method. And verify its specificity. Nine 1-day-old KM lactating mice were artificially infected. Twenty-one days later, tissues and organs such as heart, liver, spleen, lung, kidney, brain, thymus, cecum contents and blood were collected to verify the established fluorescent PCR method for testing 62 naked liver rats with cecal contents samples.

　　Result: The established detection method is highly specific. When polyoma virus is used as a template, a clear fluorescent signal will appear. When using monkey vacuole virus, mouse K virus, mouse parvovirus, rat parvovirus H-1 strain as templates, there is no fluorescent signal. The minimum detection limit of this method is 100 copies/μL. Polyoma virus nucleic acid was detected in the contents of the heart, liver, spleen, lung, kidney and cecum of artificially infected mice. The highest levels were detected in the brain, thymus and blood. The 62 samples of the cecal contents of nude mole rats were all negative for polyoma virus.

　　Conclusion: The established polyoma virus fluorescence quantitative PCR method can effectively detect polyoma virus in animal tissues. The study of nude mole mice naturally infected with polyoma virus can be used as a reference for formulating the microbiological standards of experimental nude mole mice.