动物造模 z-bio 2022-05-12 220

　　Objective: To establish a double PCR method for rapid detection of experimental rat coronavirus and Sendai virus.

　　Methods: Specific primers were designed according to rat coronavirus N gene and Sendai virus L gene; after double PCR optimization, specificity and sensitivity detection, a double PCR system was established. The PCR system was used to detect artificially infected Sendai virus tissue DNA samples and Experimental animal tissue samples were compared with ELISA method.

　　RESULTS: The target bands of rat coronavirus (168bp) and Sendai virus (262bp) were amplified by double PCR, and the PCR amplification products were sequenced using nucleic acid BLAST function to compare the homologous sequences. Sendai virus and rat coronavirus were homologous. 100% and 99%, respectively. The lower limit of detection for Sendai virus and rat coronavirus is 1.56 × 102copies/μL. The specific detection is for mouse hepatitis virus amplification, resulting in a fragment similar in size to rat coronavirus product. The PCR system detected the DNA samples of artificially infected Sendai virus tissue, and all 30 DNA samples were detected; 94 experimental animal lung tissue samples were detected, and the results were all negative.

　　CONCLUSION: The established double PCR method is simple, rapid, specific, and sensitive, and can achieve rapid detection of Sendai virus and rat coronavirus pathogens in experimental animals.