TY - JOUR N2 - The present study was aimed to establish a novel TaqMan real-time PCR (RTm-PCR) for detecting and typing bovine viral diarrhea virus (BVDV), and also to develop a diagnostic proto- col which simplifies sample collection and processing. Universal primers and TaqMan-MGB probes were designed from the known sequences of conserved 5′ - and 3′-untranslated regions (5’UTR, 3’UTR) of the NADL strain of BVDV. Prior to optimizing the assay, cDNAs were tran- scribed in vitro to make standard curves. The sensitivity, specificity and stability (reproducibility) were evaluated. The RTm-PCR was tested on the 312 feces specimens collected from persistently infected (PI) calves. The results showed the optimum conditions for RTm-PCR were 17.0 μmol/L primer, 7.5 μmol/L probe and 51.4°C annealing temperature. The established TaqMan RTm-PCR assay could specially detect BVDV without detecting any other viruses. Its detection limit was 1.55×100 copies/μL for viral RNA. It was 10000-fold higher than conventional PCR with excel- lent specificity and reproducibility. 312 samples were tested using this method and universal PCR from six dairy farms, respectively. Positive detections were found in 49 and 44 feces samples, respectively. The occurrence rate was 89.80%. In conclusion, the established TaqMan RTm-PCR could rapidly detect BVDV and effectively identify PI cattle. The detection limit of RTm-PCR was 1.55 copies/μL. It will be beneficial for enhancing diagnosis and therapy efficacy and reduce losses in cattle farms. L1 - http://czasopisma.pan.pl/Content/112846/PDF/27.pdf L2 - http://czasopisma.pan.pl/Content/112846 PY - 2019 IS - No 2 EP - 413 DO - 10.24425/pjvs.2019.129300 KW - bovine viral diarrhea virus KW - quantitative real time PCR KW - TaqMan probe A1 - Liang, H. A1 - Geng, J. A1 - Bai, S. A1 - Aimuguri, A. A1 - Gong, Z. A1 - Feng, R. A1 - Shen, X. A1 - Wei, S. PB - Polish Academy of Sciences Committee of Veterinary Sciences PB - University of Warmia and Mazury in Olsztyn VL - vol.22 DA - 2019.07.01 T1 - TaqMan real-time PCR for detecting bovine viral diarrhea virus SP - 405 UR - http://czasopisma.pan.pl/dlibra/publication/edition/112846 T2 - Polish Journal of Veterinary Sciences ER -