Development and Clinical Application of TaqMan Reverse Real Time PCR for Detection of Bovine Viral Diarrhea Virus
A TaqMan-based real-time RT-PCR assay was developed in the present study for rapid detection of bovine viral diarrhea virus(BVDV)using a pair of primers and TaqMan probe targeting the BVDV-1 conserved 5'-UTR gene sequence of domestic epidemic strains.Then this detection method was used to conduct surveillance and epidemic analysis of BVDV-1 in 7 provinces.The results showed that the developed real-time RT-PCR assay was specific for BVDV-1 and had no cross-reaction with BVDV-2,classical swine fever virus,border disease virus,bovine coronavirus,infectious bovine rhinotracheitis virus and bovine rotavirus.The detection limit of the assay was as low as 4.3 copies/μL.In addition,this assay has good reproducibility with less than 2％of the coefficient of variation within batches and between batches.Surveillance and sequence analysis of BVDV performed on large-scale cattle farms in 7 provinces showed that 17.40％samples(161/925)were positive for BVDV-1.Further phylogenetic analysis showed that BVDV-1 1a and 1c were prevalent in cattle populations.The development of a fast and sensitive diagnostic method of BVDV and findings of the dominant epidemic strain of BVDV-1a and 1c in China provided technical and data support for the early diagnosis and surveillance of BVDV.
国家科技期刊平台
NSTL
万方数据
维普
