甘蔗黄叶病毒实时荧光RT-PCR检测体系研究

Research of Real-time RT-PCR Detection System for Sugarcane Yellow Leaf Virus

甘蔗黄叶综合症(yellow leaf syndrome,YLS后称甘蔗黄叶病)是由甘蔗黄叶病毒(sugarcane yellow leaf virus,ScYLV)引起的一种重要的甘蔗病害,为更快、更准确地检测ScYLV,合成了一对特异性引物s-pf和s-pr,建立了运用SYBR Green I荧光染料法检测ScYLV的实时荧光RT-PCR体系,并对该体系的特异性、灵敏性和适用性进行了测试。结果表明:该检测体系能特异地检出ScYLV,对测试的高粱花叶病毒不能检出,灵敏度比常规PCR高100倍,适用性广。实时荧光RT-PCR检测整个过程完全闭管,无需PCR后处理且SYBR Green I荧光染料成本较低,适用于检测甘蔗体内含量较低的ScYLV病毒。

Yellow leaf syndrome caused by ScYLV （Sugarcane yellow leaf virus） is an economically important systemic a sugarcane disease in the world. So it is essential to develop a rapid and reliable method to detect ScYLV using a pair of primers s-pf and s-pr. A real-time RT-PCR system based on SYBR Green I dye has been established to detect ScYLV. The specificity sensitivity and applicability of this system were detected. The results showed that only ScYLV could be detected while sorghum mosaic virus （SrMV）cannot be detected. The sensitivity of real-time PCR is higher than that of conventional PCR by 100 fold,Real-time RT-PCR doesn＇t need post-PCR handling of the amplified products such as eleetrophoresis staining and imaging by completing the whole process within a single tube and SYBR Green I dye needs low cost so it is suitable to detect ScYLV.