动物造模 z-bio 2022-06-30 353

　　Objective: To establish a double RT-PCR method for the detection of bovine viral diarrhea virus (BVDV) in bovine samples.

　　Methods: The published BVDV type 1 and BVDV type 2 genes containing highly conserved regions in the 5'UTR region were selected as target genes, and synthetic primers were designed respectively to establish a double BVDV RT-PCR method, and the specificity, sensitivity and stability of the method were evaluated. methodological evaluation. At the same time, 41 batches of bovine-derived samples and 64 bovine plasma samples were detected by the established RT-PCR method. Bovine origin samples and 64 bovine clinical samples.

　　Results: The established BVDV RT-PCR method had no cross-reaction with bovine parainfluenza virus type Ⅲ (BPIV3), swine fever virus (CSFV) and Japanese encephalitis virus (JEV). The detection of BVDV type 1 and BVDV type 2 DNA The lowest concentration of template was 8.87×102 copies and 6.31×102 copies per microliter, respectively. BVDV type 1 and type 2 cDNAs could still detect the target band after 12 months in -30℃ refrigerator. Using the established BVDV RT-PCR method to detect 41 batches of bovine samples and 64 bovine plasma samples, the nucleic acid positive rates were 14.6% and 29.7%, respectively.

　　Conclusion: The established BVDV RT-PCR detection method has the characteristics of rapidity, specificity, sensitivity and stability, and can be used for the detection of BVDV nucleic acid in bovine samples.