动物造模,荧光定量PCR检测,诺如病毒 z-bio 2021-05-20 637

　　Objective: To establish a real-time fluorescent quantitative PCR detection method for murine norovirus, and to provide a basis for establishing a standardized detection method for MNV.

　　Method: Design specific primers based on the nucleic acid sequences of 60 MNV strains published by NCBI, construct standard positive quality control products, establish MNV real-time fluorescent quantitative PCR method and specificity, evaluate sensitivity, reproducibility and stability. This method is used to detect the cecal content samples of 766 mice under test, and to get a preliminary understanding of the MNV infection status of the Beijing experimental mice.

　　Result: We have successfully established a real-time fluorescent quantitative PCR detection method for MNV. This method is highly specific and does not cross-react with the same species of human norovirus (HuNoV) and feline calicivirus (FCV). Detection sensitivity. It can reach 101 copies/μL. The coefficient of variation of CT values within and between batches is less than 2%. Using this method, 301 MNV nucleic acid positive samples were detected in 766 mice with cecal content samples, and the positive rate was 39.3%.

　　Conclusion: The established MNV real-time fluorescent quantitative PCR detection method has excellent specificity, high sensitivity and good reproducibility, and can be used for rapid quantitative detection of MNV.