牛病毒性腹泻病病毒荧光定量PCR检测体系的建立与评价  

Establishment and evaluation of real-time PCR for detection of bovine viral diarrhea virus

作　　者：史利军[1,2] 孙宇[2] 尹惠琼[2] 孙卫华[2] 吕茂民[2] 章金刚[1,2] 

机构地区：[1]华中农业大学动物科学与动物医学院,湖北武汉430070 [2]军事医学科学院野战输血研究所,北京100850

出　　处：《中国兽医学报》2009年第12期1544-1546,共3页Chinese Journal of Veterinary Science

基　　金：全军医药卫生科研基金资助项目(06H056)

摘　　要：基于实时荧光定量PCR技术建立了一种有效地检测牛病毒性腹泻病病毒(Bovine viral diarrhea virus,BVDV)核酸的方法。对BVDV基因组进行同源比对,选取5′UTR区作为扩增目的区,经软件分析后设计特异扩增引物,扩增片段长度为203 bp。选用SYBR染料作为扩增时信号指示剂,经扩增曲线分析表明,建立的方法可有效地检测BVDV。检测体系可检测到102copies/μL的样品拷贝数。故本研究建立的BVDV实时定量检测体系可用于易感动物、牛源血液生物制品及其他可能感染或污染BVDV样品的检测。Rapid, sensitive and specific diagnostic method is necessary to confirm infection of bovine viral diarrhea virus（BVDV）. Real time PCR was used as a diagnostic method to detect and differentiate pestiviruse. In this study, a rapid real-time polymerase chain reaction for detection of BVDV was established. Primers were designed according to 5′-UTR of BVDV gene with primer prenier5. 0. In response, we developed and validated a less expensive assay with the intercalating dye SYBR green. Amplifying curve showed that this method could successfully amplify BVDV. The assay method was able to detect 10^2 copies/μL of virus genes, with correlation coefficient values of 0. 998. The newly-built real time PCR would be used for assaying BVDV in susceptible animals and blood products.

关 键 词：牛病毒性腹泻病毒 实时荧光定量PCR 5′-UTR区基因 

分 类 号：S852.65[农业科学—基础兽医学]