动物造模,多瘤病毒荧光PCR检测方法 z-bio 2021-12-16 383

　　OBJECTIVE: To establish a fluorescent quantitative PCR detection method for polyoma virus and to investigate the situation of polyoma virus infection in naked mole mice.

　　Method: 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 and specificity of the method Sex. 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 content, blood, etc. were collected to verify the established fluorescent PCR method to detect 62 samples of cecum content in naked liver rats.

　　Result: The established detection method has strong specificity, using polyoma virus as a template, and a clear fluorescence signal appears. When the monkey vacuolar virus, mouse K virus, mouse parvovirus, and rat parvovirus H-1 strain are used 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 are detected in the brain, thymus and blood. 62 samples of the cecal contents of naked mole rats were negative for polyomavirus.

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