多重RT-PCR法对马铃薯S病毒的检测效果

Detection of Potato Virus S by Multiplex RT-PCR

为探讨可信度高、特异性强、灵敏度高的马铃薯S病毒检测方法,应用RT-PCR技术建立了多重检测马铃薯S病毒的体系,并在检测过程中引入了内源性参考基因细胞色素氧化酶基因(COX)。结果表明:多重RT-PCR法灵敏度高,达128.6pg,比ELISA方法高10～100倍。多重RT-PCR检测的阳性结果与ELISA检测结果相重合,并具有更高的阳性检出率,同时还可有效避免假阴性的出现。结论:该技术检测的灵敏度高,特异性强,可信度高,可用于马铃薯S病毒检测。

In order to find a better method for detecting potato virus S with high reliability,sensitivity and strong specificity,in this study,the RT-PCR technique was used to establish the multiplex detection system of potato virus S.The potato gene Cox was used as a internal control to eliminate false negatives in the detection procedure.The result showed that the multiplex RT-PCR had high sensitivity,128.6 pg.Sensitivity test showed that RT-PCR was 10~100 times more sensitive than ELISA.The detection positive result of Multiplex RT-PCR coincided with ELISA.The multiplex RT-PCR had higher positive detection rate.Moreover,false negative was avoided effectively.This detection system is of high sensitivity,more specificity and high reliability,so it can be used for detection of potato virus S.