摘要
Abstract
To identify and differentiate rapidly the causing pathogens of clinical diseases,a multiple RT-PCR was developed.The polymerase chain reaction was optimized to simultaneously detect three pathogens,including bovine viral diarrhea virus (BVDV),bovine respiratory syncytial virus (BRSV) and bovine parainfluenza virus type 3 (BPIV-3).Three sets of specific primers were designed according to the sequences of BVDV,BRSV and BPIV-3 at the GenBank.Using three pairs of specific primers,a RT-PCR was established to amplify the conservative regions of the three viruses,respectively.The RT-PCR system would amplify a 466 bp fragment for BVDV,a 735 bp for BRSV and a 258 bp for BPIV-3 simultaneously or separately in the samples,depending on its infection status.But no specific band was amplified from other four viruses.As little as 10 pg of BVDV,1 pg of BPIV-3 and 10 pg of BRSV RNA could be detected using gel electrophoresis.Using the RT-PCR,37clinical samples were detected.The data showed that the multiple RT-PCR method was 100％ coincident with the single RT-PCR,and it can be used for the detection and differential diagnosis of three viruses.关键词
三重聚合酶链式反应/牛病毒性腹泻病毒(BVDV)/牛呼吸道合胞体病毒(BRSV)/牛副流感病毒3型(BPIV-3)Key words
multiple RT-PCR/bovine viral diarrhea virus (BVDV)/bovine respiratory syncytial virus (BRSV)/bovine parainfluenza virus type 3 (BPIV-3)分类
农业科技
