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| TaqMan荧光定量PCR检测鸡产蛋下降综合征病毒方法的建立及应用 | |
| 引用本文： | 马震原,李刚,李文超,郭宇飞.TaqMan荧光定量PCR检测鸡产蛋下降综合征病毒方法的建立及应用[J].畜牧兽医学报,2012,43(5):767-772. |
| 作者姓名： | 马震原 李刚 李文超 郭宇飞 |
| 作者单位： | 1. 四川农业大学动物医学院,雅安625014;中国农业科学院北京畜牧兽医研究所农业部兽用药物与兽医生物技术北京科学观测实验站,北京100193 2. 中国农业科学院北京畜牧兽医研究所农业部兽用药物与兽医生物技术北京科学观测实验站,北京,100193 3. 四川农业大学动物医学院,雅安625014;金陵科技学院动物科学院,南京210038 |
| 基金项目： | 国家自然科学基金，国家公益行业项目，国家高技术研究发展计划(863)项目，FAO/IAEA项目 |
| 摘 要： | 根据GenBank公布的鸡产蛋下降综合征病毒六邻体蛋白基因的高度保守序列,设计了2对特异性引物和1条TaqMan探针.以构建的阳性重组质粒为标准品,绘制标准曲线,建立了一种快速检测鸡产蛋下降综合征病毒的TaqMan荧光实时定量PCR方法.该方法最小检出量达10 copies·μL-1,在1.0×102～1.0×108copies· μL-1检测范围间有良好的线性关系,特异性、稳定性和重复性也较好.用建立的本方法检测感染鸡产蛋下降综合征病毒鸡群的产蛋分离物,与普通PCR相比,该荧光实时定量PCR方法具有更高的敏感性,可更好地用于鸡产蛋下降综合征病毒的临床检测. |
| 关 键 词： | 鸡产蛋下降综合征病毒 TaqMan探针 荧光实时定量PCR 临床检测 |
Development and Application of TaqMan Fluorescent Real-time Quantitative PCR for the Detection of Egg Drop Syndrome Virus |
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| MA Zhen-yuan , LI Gang , Li Wen-chao , GUO Yu-fei.Development and Application of TaqMan Fluorescent Real-time Quantitative PCR for the Detection of Egg Drop Syndrome Virus[J].Acta Veterinaria et Zootechnica Sinica,2012,43(5):767-772. | |
| Authors: | MA Zhen-yuan LI Gang Li Wen-chao GUO Yu-fei |
| Institution: | 1,3 (1.College of Animal Science and Veterinary Medicine,Sichuan Agricultural University,Ya’an 625014,China;2.Institute of Animal Science,Chinese Academy of Agricultural Sciences,Beijing 100093,China;3.College of Animal Science and Veterinary Medicine,Jinling Institute of Technology,Nanjing 210038,China) |
| Abstract: | To establish the fluorescent real-time quantitative PCR(FQ-RT-PCR) assay for rapid detection of egg drop syndrome virus,a pair of primers and a TaqMan probe were designed according to the sequences from highly conserved regions of the hexon protein gene of egg drop syndrome virus(EDSV).A positive recombinant plasmid was employed as standard template for the construction of standard curve.The detection limit of the assay was 10 copies DNA·μL-1 and the FQ-RT-PCR was reproducible,as shown by satisfying its wide dynamic range from 101 to 108 copies DNA·μL-1.This assay is specific and has no cross-reaction with DNA of other avian virus.The quantity of EDSV in eggs after artificial infection with NE4 strain of EDSV were detected by using the developed FQ-RT-PCR method.Compared with common PCR assay,the FQ-RT-PCR method is more sensitive and more suitable for detection of egg drop syndrome virus in vivo and in vitro. |
| Keywords: | egg drop syndrome virus TaqMan probe fluorescent real-time quantitative PCR clinical testing |
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