【摘要】 针对猪传染性胃肠炎病毒(TGEV)S基因的保守序列设计引物,以TEGV疫苗毒株为模板,克隆S基因,并构建重组阳性质粒,优化反应体系和扩增条件,建立基于SYBR GreenⅠ染料的实时荧光定量PCR检测方法,并对其特异性、重复性和灵敏度进行分析。结果显示:建立的检测方法灵敏度可达10拷贝/μL,标准曲线线性关系良好(r=0.997),扩增效率高,具有良好的特异性和重复性。使用该方法对124份临床疑似TGEV病料进行检测,阳性样本有17份,检测样本的阳性率为13.7%。该方法可用于兽医临床上TGEV的快速检测和流行病学分析。更多还原

【Abstract】 In this study,a real-time PCR method was developed for TGEV detection using SYBR Green I dye. The primers were designed according to the conserved region of TGEV S gene. The specific sequence was cloned using the attenuated TGEV strain as a template. The positive recombinant plasmid was constructed. The amplification system and conditions of real time PCR was optimized. Then the sensitivity,specificity and repeatability of this assay were analyzed. The results showed that the sensitivity of this assay was 10 copies/μL of c DNA plasmid,the correlation co-efficient of the standard curve was 0. 997. A total of 124 clinical samples were tested by this method,with 17 positive results. The positive rate of TGEV was 13. 7%. In conclusion,this method provided a technical support for laboratory diagnostic and epidemiological survey of TGEV.更多还原