| 牛病毒性腹泻病毒实时定量RT-PCR检测技术的建立及初步应用 |
| |
| 引用本文： | 任艳,陈创夫,乔军,王鹏雁,盛金良,王远志,张沾,李臻.牛病毒性腹泻病毒实时定量RT-PCR检测技术的建立及初步应用[J].畜牧兽医杂志,2010,29(5):4-7. |
| |
| 作者姓名： | 任艳 陈创夫 乔军 王鹏雁 盛金良 王远志 张沾 李臻 |
| |
| 作者单位： | 1. 石河子大学动物科技学院,新疆,石河子832003
2. 石河子大学医学院 |
| |
| 摘 要： | 以牛病毒性腹泻病毒(BVDV)的保守基因5'-非编码区为参考,设计、优化一对特异的荧光定量PCR引物,应用Smart CycleⅡ分析系统,结合Eva Green荧光染料结合原理,建立一种快速、定量检测牛病毒性腹泻病毒的荧光定量PCR技术。该方法线形范围为101~106copies/μL,相关系数为R2=0.997,扩增效率为E=0.78。灵敏性比常规PCR高102倍。该方法具有良好的特异性,检测变异系数低于1%。应用本实验建立的方法检测25份不同地区采集的临床症状疑似BVDV感染的牛粪样品,阳性检出率为72%(18/25),与病毒分离培养和电检观察相比,该方法具有快速、灵敏、特异、重复性好和能定量检测等优点,为实验室快速、准确检测BVDV奠定了基础。
|
| 关 键 词： | 牛病毒性腹泻病毒 实时定量RT-PCR 检测 |
Establishment and Initial Application of a Real-time Fluorescent Quantitative PCR Method for Detection of Bovine Viral Diarrhea Virus |
| |
| REN Yan,CHEN Chuang-fu,QIAO Jun,WANG Peng-yan,SHENG Jin-liang,WANG Yuan-zhi,ZHANG Zhan,LI Zhen.Establishment and Initial Application of a Real-time Fluorescent Quantitative PCR Method for Detection of Bovine Viral Diarrhea Virus[J].Journal of Animal Science and Veterinary Medicine,2010,29(5):4-7. |
| |
| Authors: | REN Yan CHEN Chuang-fu QIAO Jun WANG Peng-yan SHENG Jin-liang WANG Yuan-zhi ZHANG Zhan LI Zhen |
| |
| Institution: | REN Yan1,CHEN Chuang-fu1 QIAO Jun1,WANG Peng-yan1,SHENG Jin-liang1,WANG Yuan-zhi2,ZHANG Zhan1,LI Zhen1 (1.College of Animal Science and Technology,Shihezi University,Shihezi Xinjiang 832003,China,2.College of Medicine,Shihezi University) |
| |
| Abstract: | The aim of this study was to develop a Eva-Green real-time quantitative PCR which can detect Bovine viral diarrhea virus(BVDV) quickly and flexibly.According to genome sequences within 5'-UTR of BVDV published in GenBank,a pair of primers was designed.The assay had a dependent coefficient R2=0.997,amplified efficiency E=0.78 and detection sensibility is 100 times than conventional PCR,with variation coefficient less than 1.0%.In this study,twenty five samples from bovine were detected by this method.Eightee... |
| |
| Keywords: | Bovine viral diarrhea virus fluorescent quantitative RT-PCR establishment |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ |
|
