动物造模,牛冠状病毒,荧光定量PCR z-bio 2021-06-25 261

　　Objective: To establish a specific and sensitive fluorescent quantitative PCR detection method for bovine coronavirus, and to screen cattle-derived samples.

　　Methods: According to the N gene sequence of BCV registered in Genbank, TaqMan primer probes were designed, and a BCV PCR detection method based on TaqMan probe method was established. The collected bovine samples were screened for BCV, and the PCR products of some positive samples were tested. Sequencing identification.

　　Results: A TaqMan probe fluorescence quantitative PCR detection method that can specifically detect BCV was established. The standard curve correlation coefficient Slope was -3.417, the correlation coefficient r2 was 0.999, and the amplification efficiency Eff was 96.195%. This method is sensitive to the detection of BCV. The detection rate is 40 copies. Using this method, the detection rate of 64 bovine anal swab samples was 1.5%, and the detection rate of 33 bovine biological products was 6%. The PCR products of some positive samples were compared to BCV nucleic acid by sequencing. The sequence indicates that BCV is prevalent in domestic cattle, and there is a potential risk of BCV infection in biological products of cattle origin.

　　Conclusion: After sequencing verification, the established method can sensitively detect the BCV nucleic acid in the sample. The monitoring of BCV in biological products related to cattle should be strengthened to avoid the potential risk of human infection with BCV.