鸡传染性支气管炎病毒荧光定量PCR标准曲线的建立

Construction of Standard Curve for Real-time PCRAssay of Infectious Bronchitis Virus

笔者针对传染性支气管炎病毒(IBV)M41株中编码核衣壳蛋白的N基因设计并合成一对引物,通过构建重组阳性标准质粒的方法,成功建立SYBR GreenⅠ荧光定量PCR方法检测IBV的标准曲线。该方法所建立的曲线可检测到初始模板中5×102copies/μL的病毒核酸,与常规PCR相比敏感性大大提高。笔者建立的荧光定量PCR检测方法具有灵敏度高、特异性强、重复性好等特点,可用于传染性支气管炎的临床诊断和健康带毒鸡群的检测。

In this study, the authors constructed standard curve of Real-time PCR assay for detecting infectious bronchitis virus（IBV）. In this experiment, the dye of SYBR Green I was used. A pair of primers was designed according to the sequences of the N gene which coding nucleocapsid protein of M41strain. And authors used positive recombinant plasmid as standard to construct the standard curve. This method had a detection limit of 5 × 10^2 copies/μL viral RNA, its sensitivity greatly enhanced compared to conventional PCR. The results of the test showed that the Real-time PCR could be used for the clinical diagnosis of IBV infection because of its high sensitivity, specificity and repeatability.