Mehle Nataša, Nikolić Petra, Gruden Kristina, Ravnikar Maja, Dermastia Marina
National Institute of Biology, Ljubljana, Slovenia.
Methods Mol Biol. 2013;938:269-81. doi: 10.1007/978-1-62703-089-2_23.
In this chapter, we describe a real-time PCR detection system for fast, reliable, specific, and sensitive detection and discrimination of 'Candidatus Phytoplasma mali', 'Ca. P. prunorum', and 'Ca. P. pyri' from the 16SrX (apple proliferation-AP) group. These phytoplasmas are causal agents of fruit tree diseases within the Rosaceae family, namely apple proliferation, European stone fruit yellows, and pear decline. The assays use (hydrolysis) TaqMan(®) minor groove binder probes. The panel of assays comprises the same set of primers and specific probes for species-specific amplification, and an additional set of primers and probe for 18S rRNA as an endogenous quality control of DNA extraction. The assays described can be used in routine phytoplasma surveys and in certification programmes.
在本章中，我们描述了一种实时PCR检测系统，用于从16SrX（苹果增殖-AP）组中快速、可靠、特异且灵敏地检测和鉴别“苹果植原体暂定种（‘Candidatus Phytoplasma mali’）”、“李属植原体暂定种（‘Ca. P. prunorum’）”和“梨属植原体暂定种（‘Ca. P. pyri’）”。这些植原体是蔷薇科果树病害的病原体，即苹果增殖病、欧洲核果黄化病和梨衰退病。这些检测方法使用（水解）TaqMan®小沟结合探针。该检测方法组包括用于物种特异性扩增的同一组引物和特异性探针，以及用于18S rRNA的另一组引物和探针，作为DNA提取的内源性质量控制。所描述的检测方法可用于常规植原体调查和认证计划。
