畜牧兽医学报2012，Vol.43Issue(5)：767-772,6.
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
Development and Application of TaqMan Fluorescent Real-time Quantitative PCR for the Detection of Egg Drop Syndrome Virus
摘要
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 10' 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.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Key words
egg drop syndrome virus/ TaqMan probe/ fluorescent real-time quantitative PCR/clinical testing分类
农业科技引用本文复制引用
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):767-772,6.基金项目
国家自然科学基金(31172342) （31172342）
国家公益行业项目(200903037-2) （200903037-2）
国家高技术研究发展计划(863)项目(2008AA102411) （863）
FAO/IAEA项目(14515) （14515）
