中国动物传染病学报2016，Vol.24Issue(1)：7-14,8.
一种新型鸭呼肠孤病毒SYBR Green II荧光定量PCR方法的建立
DETECTION OF NOVEL DUCK REOVIRUS USING SYBR GREEN II FLUORESCENT QUANTITATIVE PCR ASSAY
摘要
Abstract
To develop a fast, sensitive, specific SYBR Green II fluorescent quantitative PCR assay for detecting Novel duck reovirus (NDRV) infection, S1 gene was amplified in RT-PCR from the NDRV infected-duck tissues, and cloned into pET-30a vector. The resulting recombinant plasmid was used as the template for making a standard curve. Subsequently, the sensitivity and specificity of the SYBR Green II fluorescent quantitative PCR assay that was developed were determined. The results showed that the NDRV real-time PCR assay had a dynamic range of detection between 101 and 108 copies/μL with a sensitivity of 10 copies/μL. There was no cross reaction with H5 subtype Avian influenza virus, H9 subtype Avian influenza virus, Infectious bronchitis virus, Duck hepatitis virus type C, New castle disease virus, Goose parvovirus and Duck viral enteritis. In conclusion, a SYBR Green II fluorescent quantitative PCR assay has been developed for quantification of NDRV, which can be sued for investigating the pathogenesis of NDRV.关键词
新型鸭呼肠孤病毒/S1/荧光定量PCR/熔解曲线/检测Key words
Novel duck reovirus/S1/fluorescent quantitative PCR/melting curve/detection分类
农业科技引用本文复制引用
丁明洋,戚伟强,陈宗艳,朱杰,吴巧梅,缪秋红,李传峰,吴润,刘光清..一种新型鸭呼肠孤病毒SYBR Green II荧光定量PCR方法的建立[J].中国动物传染病学报,2016,24(1):7-14,8.基金项目
上海市科委创新项目（13391901600）；国家自然科学基金项目 （）
