Detection of Infectious Bursal Disease Virus Using RT-PCR SYBR GreenⅠ Technology

Conservative fragment of VP5 gene was amplified by RT-PCR and cloned into pMD18-T vector.After sequencing,the positive recombinant plasmid was acquired and used to establish standard and melt curves of the real-time fluorescent quantitative RT-PCR.The standard curve for the threshold cycle and viral genomic copy number ranging from 3.2910～3.29108 were linear.The sensitivity of detection was 33 copies.It was concluded that the real time RT-PCR SYBR GreenⅠ technology was highly specific and repeatable for IBDV diagnostics and quantification.