中国兽医科学2011，Vol.41Issue(2)：193-198,6.
鸡传染性支气管炎病毒real-time PCR检测方法的建立
Development of a real-time PCR assay for detection of infectious bronchitis virus
摘要
Abstract
A pair of primers was designed according to the sequence of the N gene fragment of infectious bronchitis virus(IBV) and a recombinant plasmid containing the target gene was constructed as a standard control.A real-time PCR assay based on SYBR Green Ⅰ was developed for detection of IBV.The developed assay had a good linear relationship, and the correlation coefficient of the standard curve was 0.998 3.The assay had a detection limit of 10 copies/μL of initial templates and had no cross-reaction with Newcastle disease virus, infectious laryngotracheitis virus, infectious bursal disease virus and Mark's disease virus,and had a coefficient of variations less than 3％ for both intra- and inter-assay.40 samples of trachea and kidney from 20 experimentally infected chickens were examined by the developed real-time PCR assay,and the results showed that the kidney had higher virus loads than the trachea on day 5 and 7 post-infection, indicating that the clinical feature were related to viral RNA loads.Therefore, this real-time PCR assay is sensitive,specific and repeatable and could be used as an effective assay for detection of nucleic acids and quantification of viral loads of IBV.关键词
传染性支气管炎病毒/实时荧光定量聚合酶链反应/SYBR Green Ⅰ分类
农业科技引用本文复制引用
磨美兰,陈秋英,侯金莲,范文胜,李孟,韦平,韦天超..鸡传染性支气管炎病毒real-time PCR检测方法的建立[J].中国兽医科学,2011,41(2):193-198,6.基金项目
国家自然科学基金项目(30700599) （30700599）
广西自然科学基金项目(桂科自 0991044) （桂科自 0991044）
广西科技攻关项目(0993009-2) （0993009-2）
广西大学拔尖创新团队建设计划项目 （）
