摘要
为建立鸭瘟病毒(DPV)快速、敏感的检测方法,本研究利用PCR技术扩增出DPV UL30基因中510bp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DPV的SYBR Green I荧光定量PCR检测方法。该方法检测灵敏度可达5×101拷贝,与鸭细小病毒、鸭圆环病毒、小鹅瘟病毒、鸭肝炎病毒、鸭H9亚型流感病毒和鸭副粘病毒均不发生交叉反应,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DPV的检测。
To develop the SYBR Green I real-time quantitative PCR assay for detection of Duck Plague virus （DPV）, a 510bp conservative region from DPV UL30 gene was amplified by RT-PCR and cloned into pMD18-T vector to construct recombinant plasmid of pMD18-UL30,which was served as template to establish the standards curve of the SYBR Green I real time PCR.The results showed that the SYBR Green I real-time PCR was specifically detected DPV with a limited detection of 5×101copies and no amplification for Duck Parvovirus, duck circovirus, gosling plague virus, duck hepatitis virus, H9 subtype duck influenza virus and Duck Paramyxovirus.There data suggested that the SYBR Green I realtime PCR could be applied in clinical diagnosis and epidemiological investigation for DPV.
基金
山东省现代产业技术体系家禽生产与环境控制创新团队项目(SDAIT-13-011-10)
