动物造模,牛病毒性腹泻病毒 z-bio 2021-10-14 428

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

　　Method: Select the published BVDV1 and BVDV2 genes containing the highly conserved 5'UTR region as target genes, design synthetic primers, and establish a double BVDVRT-PCR method to determine gender specificity, sensitivity and stability. Methodological evaluation methods, etc. At the same time, the established RT-PCR method was used to detect 41 batches of bovine samples containing bovine serum, bovine serum deproteinized extract, spleen polypeptide injection, and 64 bovine plasma samples. Source samples of dairy cows and clinical samples of 64 dairy cows.

　　Results: The established BVDVRT-PCR detection method did not cross-react with bovine parainfluenza virus type III (BPIV3), swine fever virus (CSFV), Japanese encephalitis virus (JEV), and BVDV1 and BVDV2 DNA templates. The minimum detection concentration was per micron The liters are 8.87 x 102 servings and 6.31 x 102 servings. BVDV type 1 and type 2 cDNA can detect the target band even if placed in a refrigerator at -30°C for 12 months. Using the established BVDVRT-PCR method, 41 batches of cattle samples and 64 cattle plasma samples were detected, and the nucleic acid positive rates were 14.6 29.7?

　　Conclusion: The established BVDVRT-PCR detection method is fast, specific, sensitive and stable, and can be used to detect BVDV nucleic acid in cattle samples.