华北农学报2017，Vol.32Issue(z1)：49-53,5.DOI:10.7668/hbnxb.2017.S1.009
BVDV SYBR GreenⅠ荧光定量PCR方法与RT-PCR方法的建立与应用
The Developments of SYBR Green Ⅰ Real-time PCR and Conventional RT-PCR for Detection of BVDV
摘要
Abstract
Rapid,sensitive and specific diagnostic method is necessary to confirm infection of Bovine viral diar-rhea 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 de-signed 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 GreenⅠcould 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 typeⅡBVDV strain).As for the limit of detection of per method,the limit of detection of RT-PCR could reach to 10-1TCID50/mL;the limit of detection of Real-time PCR with SYBR GreenⅠcould reach to 3.4×102 copies/μL.The two methods were used to detect clinical serum samples efficiently,suggesting that the current develop-ments of detection methods can take application for epidemiology of BVDV and clinical assay of BVDV.关键词
牛病毒性腹泻病毒(BVDV)/RT-PCR/SYBRGreenⅠ荧光定量PCR/应用Key words
Bovine viral diarrhea virus(BVDV)/RT-PCR/SYBR Green Ⅰ Real-time PCR/Application分类
农业科技引用本文复制引用
马鹏,李林杰,常秋燕,王悦萦,郭富城,刘萍,马晓霞,马忠仁..BVDV SYBR GreenⅠ荧光定量PCR方法与RT-PCR方法的建立与应用[J].华北农学报,2017,32(z1):49-53,5.基金项目
企业横向项目(No.XBMu-2015-Bc-11) （No.XBMu-2015-Bc-11）
