摘要
目的:建立枣疯病植原体拷贝数检测实时荧光定量PCR方法,为枣疯病植原体定量检测提供技术支持。方法:构建质粒标准品,设计实时荧光PCR探针引物,优化体系,建立标准曲线,并进行重复性验证。结果:制备了枣疯病植原体标准质粒,建立了稳定的质粒标准品检测体系(R2=0.998,检测限10拷贝,定量限100拷贝)。结论:实时荧光定量PCR检测方法重复性好,可用于枣疯病植原体的拷贝数检测,为枣疯病植原体检验检测和病害防治提供了技术支持。
Objective：To establish the quantitative detection method of Jujube Watches＇Broom（JWB）by realtime PCR,and provide a technical support for the JWB detection.Methods：Standard plasmid was constructed, and then a set of primers and probe was designed.Optimizing the real-time PCR system,the standard curve was created.The reproducibility was also verified.Results：The standard plasmid was constructed successfully,and a stable detection system was set up.In this detection system,the LOD is 10 copies and the LOQ is 100 copies （coefficient of determination R2=0.998）.Conclusion：This system has a sufficient repeatability that can be used as a quantitative study and also can provide technical support for the detection and prevention of JWB.
出处
《生物技术通讯》
CAS
2010年第1期70-72,85,共4页
Letters in Biotechnology
基金
国家自然科学基金(30671395)
中国检验检疫科学研究院基本科研业务专项(2008JK005)
