Hodgetts Jennifer, Dickinson Matt
The Food and Environment Research Agency, Sand Hutton, York, UK.
Methods Mol Biol. 2013;938:233-44. doi: 10.1007/978-1-62703-089-2_20.
Conventionally, detection of phytoplasmas has been performed by PCR of the 16S rRNA gene, followed by either RFLP or DNA sequencing to determine the phytoplasma 16Sr group. This chapter demonstrates the technique of terminal restriction fragment length polymorphism (T-RFLP), a fingerprinting technique which combines both detection and identification in a single method, with the added benefit of inbuilt controls which removes the risk of false negative results and in addition highlights potential false positive results.
传统上,植原体的检测是通过对16S rRNA基因进行PCR,然后进行限制性片段长度多态性分析(RFLP)或DNA测序来确定植原体的16Sr组。本章展示了末端限制性片段长度多态性(T-RFLP)技术,这是一种指纹识别技术,它将检测和鉴定结合在一种方法中,还有内置对照的额外好处,可消除假阴性结果的风险,此外还能突出潜在的假阳性结果。