动物造模,荧光PCR检测 z-bio 2022-03-14 440

　　Objective: To establish a quantitative PCR detection method for polyoma virus, and to investigate the infection of polyoma virus in naked mole rats.

　　Methods: The nucleic acid sequences of mouse polyoma virus (Genbank: NC_001515) in NCBI were compared, and the conserved regions were selected to design primers and probes, and a quantitative PCR method for polyoma virus was established to verify the sensitivity and specificity of the method. Nine 1-day-old KM suckling mice were artificially infected. After 21 days, the heart, liver, spleen, lung, kidney, brain, thymus, cecum contents, blood and other tissues and organs were collected. The established fluorescent PCR method was used to detect and verify the method. Validity in application, the method was finally used to detect 62 samples of cecal contents of naked mole rats.

　　Results: The established detection method has good specificity, and obvious fluorescence signal appeared when polyoma virus was used as the template. The simian vacuolar virus, mouse K virus, mouse parvovirus and rat parvovirus H-1 strain were used as templates. No fluorescent signal appeared; the minimum detection limit of the method was 100 copies/μL; polyoma virus nucleic acid was detected in the heart, liver, spleen, lung, kidney and cecum contents of artificially infected mice, of which the content in lung tissue Highest, undetectable in brain, thymus, and blood; 62 samples of caecal contents of naked mole rats tested negative for polyomavirus.

　　Conclusion: The established polyomavirus real-time PCR method can effectively detect polyomavirus in animal tissues, and the investigation of natural infection of naked mole-rats with polyomavirus provides a reference for the formulation of microbial standards for experimental naked mole-rats.