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.
出处
《中国兽医学报》
CAS
CSCD
北大核心
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)
