Abstract: Rapid,sensitive and specific diagnostic method is necessary to confirm infection of Bovine viral diarrhea virus(BVDV).RT-PCR and Real-time PCR with SYBR Green Ⅰ was used as a diagnostic method to detect BVDV.According to genomes of Bovine viral diarrhea virus(BVDV) in GenBank,a pair of primers had been designed to develop conventional RT-PCR method and Real-time PCR with SYBR Green Ⅰ for detection of BVDV.Depending on estimations of specificity and sensitivity and stability of the two methods,RT-PCR method could amplify the specific PCR product with 250 bp in length from BVDV standard strain (Oregon C24),and Real-time PCR with SYBR Green Ⅰ could also obtain positive signal.The two methods didn't obtain any signals from BPV,BPIV3,BAV-3 and REO which are main pathogens for cattle industry,but could obtain positive signals from some BVDV strains (C24V,GSTZ,Av69,S183 and type Ⅱ BVDV strain).As for the limit of detection of per method,the limit of detection of RT-PCR could reach to 10^-1 TCID₅₀/mL;the limit of detection of Real-time PCR with SYBR Green Ⅰ could reach to 3.4×10² copies/μL.The two methods were used to detect clinical serum samples efficiently,suggesting that the current developments of detection methods can take application for epidemiology of BVDV and clinical assay of BVDV.

Key words: Bovine viral diarrhea virus(BVDV), RT-PCR, SYBR Green Ⅰ Real-time PCR, Application

CLC Number: 

• S432.4