【摘要】 为建立一种特异、灵敏、量化、快速的猪流行性腹泻病毒(PEDV)检测方法,设计1对PEDV N基因的特异性引物,建立Eva Green染料的实时荧光定量RT-PCR的检测方法,并对疑似PEDV感染的临床样品进行检测。结果显示,所建立的实时荧光定量RT-PCR方法的标准曲线具有良好的线性关系,线性相关系数R2=0.999;不与猪传染性胃肠炎病毒(TGEV)、猪瘟病毒(CSFV)、猪繁殖与呼吸综合征病毒(PRRSV)发生交叉反应,具有较强的特异性;灵敏度比常规RT-PCR方法高100倍;重复性试验的变异系数均小于5%,重复性良好。对43份临床样品进行检测,结果比普通PCR多检出2份阳性。结果表明,建立的实时荧光定量RT-PCR检测方法在特异性、灵敏度和重复性方面都很好,可用于临床PEDV检测。更多还原

【Abstract】 To establish a quick,specific,sensitive and quantitative detection method of porcine epidemic diarrhea virus(PEDV),apair of specific primers were designed on the basis of conservative sequence of PEDV N gene.Eva Green RT-qPCR was established,and its specificity,sensitivity,repeatability were determined.The results showed that the standard curve of RT-qPCR revealed a good linear relationship with the correlation coefficient R~2=0.999,high specificity that no cross-reactions with TGEV,CSFV and PRRSV,high sensitivity that 100 times more sensitive than the conventional RT-PCR method,and well repeatability with the coefficient of variation less than 5%.This method had high specificity,good sensitivity and good repeatability,and can be used for clinical detection of PEDV.更多还原