摘要
为了建立禽呼肠孤病毒(ARV)快速、敏感的检测方法。本研究利用RT-PCR技术扩增出ARV S1基因中298bp的保守序列,并克隆到p MD18-T载体中作为标准品制作标准曲线,建立了ARV的SYBR Green Ⅰ荧光定量PCR检测方法。该方法检测灵敏度可达4.8×101拷贝,与NDV、AIV、IBV、IBDV、ALV、REV均不发生交叉反应,具有良好的特异性和重复性。结果表明,建立的实时荧光定量PCR具有特异、敏感、快速、定量、重复性好等优点,可用于临床ARV的检测。
To develop the SYBR Green Ⅰ real-time quantitative PCR assay for detection of Avian reovirus（ARV）, a 298 bp conservative region from ARV S1 gene was amplified by RTPCR and cloned into p MD18-T vector to construct recombinant plasmid of pMD18-S1,which was served as template to establish the standards curve of the SYBR Green Ⅰ rea1 time PCR.The results showed that the SYBR Green I real-time PCR was specifically detected ARV with a limited detection of 4.8 ×10^1copies and no amplification for NDV 、AIV 、IBV 、IBDV 、ALV and REV. There data suggested that the SYBR Green I real-time PCR could be applied in clinical diagnosis and epidemiological investigation for ARV.
出处
《家禽科学》
2015年第10期10-14,共5页
Poultry Science
基金
山东省现代产业技术体系家禽创新团队生产与环境控制创新团队项目(SDAIT-13-011-10)
