中国畜牧兽医2012，Vol.39Issue(4)：169-173,5.
鸡传染性支气管炎病毒H52株TaqMan-MGB荧光定量RT-PCR检测方法的建立
Establishment of TaqMan-MGB Fluorescence Quantitative RT-PCR Assay for Detection of Avian Infectious Bronchitis Virus H52 Strain
摘要
Abstract
A rapid assay for detecting avian infectious bronchitis virus was established and laid the foundation for further study on new methods of vaccine potency testing and quality control. Primers and probe of fluorescence quantitative PCR were designed for H52 strain, according to the N gene of IBV. Then the method was optimized. Specificity, sensitivity, and repeatability were tested and the correlation was explored between the quantitative RT-PCR method and EID50 on the determination of virus quntity. In addition, in vitro transcription technology was used to develop a quantitative PCR format with virus copies amount. It was highly specific and no cross-reactions were found with other avian pathogens. The assay had a detection limit of 5. 60×101 copies/μL viral RNA, which was 10 times more sensitive than the conventional PCR. Coefficient of variation value ranged from 0. 9% to 3. 2% in repeatability test. Compared with EID50 test results, the correlation coefficient r was 0. 934, which showed that two methods in detecting the virus titer on the H52 strain had a good correlation. This assay of TaqMan-MGB fluorescence quantitative RT-PCR is suitable for rapid and quantitative detection of IBV H52 strain.关键词
荧光定量RT-PCR/TaqMan-MGB探针/鸡传染性支气管炎病毒/H52Key words
luorescence quantitative RT-PCR/TaqMan-MGB probe/avian infectious bronchitis virus/H52分类
农业科技引用本文复制引用
梁耀峰,郭霄峰,宋立,刘洋..鸡传染性支气管炎病毒H52株TaqMan-MGB荧光定量RT-PCR检测方法的建立[J].中国畜牧兽医,2012,39(4):169-173,5.基金项目
“十一五”国家支撑计划子课题(2006BAK02A03-1). （2006BAK02A03-1）
