摘要
RT-PCR扩增H9亚型禽流感病毒(AIV(H9))的HA基因保守片段,将其克隆到pMD18-T载体,经测序验证正确作为阳性质粒。以阳性质粒为标准品,建立SYBR Green I荧光定量RT-PCR的标准曲线。结果表明,该方法灵敏性可达74.3copies/μL,且特异性好、重复性佳。SYBR Green I实时荧光定量RT-PCR方法为H9亚型禽流感的病原的快速诊断和病毒的定量分析提供重要手段。
Conservative fragment of HA gene of AIV（H9） was amplified by RT-PCR and cloned into pMD18-T vector.After sequencing,the positive recombinant plasmid was acquired and used to establish standard curve of the SYBR Green Ⅰ real-time RT-PCR.The results showed that the sensitivity was so good that 74.3 copies/μL of the virus content could be detected.Furthermore,this method was highly specific and reproducible.It is concluded that the SYBR Green Ⅰ real-time RT-PCR is a good technology for pathogenic diagnosis and virus quantification of AIV（H9）.
出处
《青岛农业大学学报（自然科学版）》
2013年第3期195-199,共5页
Journal of Qingdao Agricultural University(Natural Science)
基金
青岛市公共领域科技支撑计划(10-3-3-17-nsh)
山东省现代农业产业技术体系家禽创新团队项目
作者简介
徐守振（1977-）,男,山东沂水人,副教授,博士,主要从事禽病防治研究.
