| 伪狂犬病毒荧光定量PCR检测方法的建立 |
| |
| 引用本文： | 田云,任裕其,屈源泉,孙彦伟,卢受异,卢洪芬,李明,孔令辰,王连想.伪狂犬病毒荧光定量PCR检测方法的建立[J].广东畜牧兽医科技,2009,34(4):31-33. |
| |
| 作者姓名： | 田云 任裕其 屈源泉 孙彦伟 卢受异 卢洪芬 李明 孔令辰 王连想 |
| |
| 作者单位： | 广东省动物防疫监督总所,广东,广州,510230 |
| |
| 摘 要： | 根据猪伪狂犬病毒gD基因的序列，设计和合成了一对特异的可用于检测伪狂犬病毒的PeR引物和一条Taqman荧光探针，采用LightCycle 480荧光定量PCR仪，建立了一种可实时定量检测猪伪狂犬病毒的荧光定量PCR技术。该方法的线性范围为1．0×10^1-1．0×10^10拷贝，灵敏度可达2拷贝，比常规PCR高100倍。该方法检测时间短，速度快，仪器的运行时间仅为1h，特异性明显高于常规PCR。对15株猪伪狂犬病毒进行了检测，结果均为阳性；与猪细小病毒和鸭瘟病毒无非特异性反应。与病毒分离培养和常规PeR相比较，该方法具有快速、灵敏、特异、定量、重复性好的优点，可望用于，临床上伪狂犬病的检测和病毒分布的研究等。
|
| 关 键 词： | 猪伪狂犬病毒 荧光定量PCR 临床检测 |
Establishment of a quantitative real-time PCR method for detection of pseudorabies virus |
| |
| Tian Yun,Ren Yuqi,Qu Yuanquan,Sun Yanwei,Lu Shousheng,Lu Hongfen,Li Ming,Kong Lingchen,Wang Lianxiang.Establishment of a quantitative real-time PCR method for detection of pseudorabies virus[J].Guangdong Journal of Animal and Veterinary Science,2009,34(4):31-33. |
| |
| Authors: | Tian Yun Ren Yuqi Qu Yuanquan Sun Yanwei Lu Shousheng Lu Hongfen Li Ming Kong Lingchen Wang Lianxiang |
| |
| Institution: | (Guangdong Animal Health Supervision Institute, Guangzhou 510230,China) |
| |
| Abstract: | According to the sequences of the gD gene ofpseudorabies virus（PRV）, one pairs of primers and one Taqman probe were designed and synthesized for detection of PRV. By using LightCycler 480 real-time PCR machine, a real-time and quantitative PCR method was established. The dynamic range of the quantitative real-time PCR was from 1.0×10^1to1.0×10^10 copies and the lowest DNA detection limit was 2 copies, which was 100 fold better than regular PCR. The detection method was rapid and only one hour was needed for detection on PCR machine. It was more specific than regular PCR. Fifteen pseudorabies virus strains were detected by this PCR method and all were positive. There were no amplification with swine parvovirus and duck plague virus. Compared with virus isolation and regular PCR, this method was more rapid, sensitive, specific and repeatable. It can be used on the detection for PRV and the research on tissue distribution of PRV, etc. |
| |
| Keywords: | Pseudorabiesvirus（PRV） quantitativereal-timePCR clinicaldetection |
| 本文献已被 维普 万方数据 等数据库收录！ |
|
