利用内标为基础的RT-PCR技术检测草莓皱缩病毒(SCV)和草莓镶脉病毒(SVBV)

Detection of SCV and SVBV with Multiplex RT-PCR Based on Internal Control

[目的]通过对反应条件和参数的摸索与优化,建立多重RT-PCR检测方法,实现2种病毒的同时高效快速检测。[方法]通过CTAB法与试剂盒提取RNA,以优质的总RNA为模板,进行梯度PCR,结合扩增内标的检测体系,建立多重RT-PCR检测方法。[结果]建立了优化的SCV和SVBV的多重RT-PCR检测体系,可以对草莓田间植株进行快速稳定的病毒检测。[结论]该研究为草莓病毒检测提供一种方便、高效的分子生物学方法,为草莓无病毒苗的实际生产提供技术支撑。

[ Objective ] Through optimization of reaction conditions and parameters, multiple RT-PCR detection method was established in order to realize efficient and rapid detection of two kinds of virus. [ Method] By CTAB method and the kit, RNA was extracted. Using highquality total RNA as a template, gradient PCR was done, Combining with internal amplification target detection system, a multiplex RT-PCR detection system of SCV and SVBV was established. [ Result] A multiplex RT-PCR detection system of SCV and SVBV was established, and could make fast and stable virus detection in field grown strawberries. [ Couclusion] The study provided a convenient and efficient molecular biology method for strawberry virus detection, and provided technical support for actual production of strawberry virus-free seedling.