摘要
RT-PCR扩增IBDV的VP5基因保守片段,将其克隆到pMD18-T载体,经测序鉴定得到阳性质粒。以阳性质粒为标准品,建立SYBR GreenⅠ实时荧光定量RT-PCR的标准曲线和溶解曲线。结果表明,IBDV荧光定量PCR的标准曲线Ct值与3.29×10~3.29×108之间的病毒基因拷贝数呈现良好的线性关系。该方法灵敏度可达33拷贝,且特异性和重复性好。SYBR GreenⅠ实时荧光定量RT-PCR方法可用于IBDV的病原诊断和病毒的定量分析。
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 3.29×10～3.29×108 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.
出处
《福建农业学报》
CAS
2011年第2期175-179,共5页
Fujian Journal of Agricultural Sciences
作者简介
作者简介：朱春华（1980-），女，助理研究员，硕士，主要从事动物病毒学研究（E-mail：zchixd80@163．com）
通讯作者：黄瑜（1965-），男，研究员，博士，主要从事动物传染病研究（E—mail：huangyu_815@163．com）
