运用实时荧光RT-PCR技术检测柑橘碎叶病毒  

Detection of Citrus tatter leaf virus(CTLV) by real-time RT-PCR

作　　者：刘科宏[1] 周常勇[1] 宋震[1] 周彦[1] 李中安[1] 唐科志[1] 

机构地区：[1]西南大学柑橘研究所,重庆400712

出　　处：《果树学报》2009年第5期748-751,共4页Journal of Fruit Science

基　　金：国家支撑计划(2007BAD47B03;2007BAD61B04);重庆市自然科学基金(2007BBB44)

摘　　要：柑橘碎叶病是由橘碎叶病毒(Citrus tatter leaf virus,CTLV)引起的一种重要的柑橘病害,为更快、更准确的检测CTLV,合成了一对特异性引物ASG-Pf和ASG-Pr,建立了运用SYBRGreenI荧光染料法检测CTLV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,该检测体系能特异的检出CTLV,对测试的衰退病毒、温州蜜柑萎缩病毒和鳞皮病毒都不能检出;灵敏度比常规PCR高100倍;适用性广,可检测出多种柑橘类植物中的CTLV。实时荧光RT-PCR检测整个过程完全闭管,无需PCR后处理,且SYBR GreenI荧光染料成本较低,适用于检测柑橘体内含量较低的CTLV病毒。Citrus tatter leaf disease caused by citrus tatter leaf virus （CTLV） is an economically important systemic disease of citrus in the world. So it is essential to develop a rapid and reliable method to detect CTLV. Using a pair of primers ASG-Pf and ASG-Pr, a real-time RT-PCR system based on SYBR Green I dye has been established to detect CTLV. The specificity, sensitivity and applicability of this system were detected. The results showed that only CTLV could be detected,while Citrus tristeza virus （CTV）, Satsuma dwarf virus （SDV）and Citrus psorosis virus （CPV） can not be detected. The sensitivity of real-time PCR is higher than that of conventional PCR by 100 fold,and it can detect CTLV isolates from different kinds of citrus cultivars. Real-time RT-PCR doesn＇t need post-PCR handling of the amplified products such as electrophoresis, 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 CTLV.

关 键 词：柑橘碎叶病毒(CTLV) 实时荧光RT—PCR 检测 

分 类 号：S666[农业科学—果树学]