Rural Economy & Veterinary Services, Regional Sector of Piraeus, Attica Prefecture, Piraeus, Greece.
Department of Crop Science, Agricultural University of Athens, Athens, Greece.
Methods Mol Biol. 2022;2316:195-210. doi: 10.1007/978-1-0716-1464-8_17.
A wide range of nucleic acid-based detection methods is available for viroid diagnosis. In this chapter, we describe a novel reverse transcription loop-mediated isothermal amplification (RT-LAMP). The technique involves the use of nucleic acids, plant sap, or even traces of plant tissue. Adding a set of four to six primers, Bst DNA polymerase, AMV, or another thermostable reverse transcriptase, allows the fast detection of RNA targets like viroids after a short (20-40 min) incubation at 63 °C either in an open or close-tube system.
有多种基于核酸的检测方法可用于类病毒诊断。在本章中，我们描述了一种新型的逆转录环介导等温扩增（RT-LAMP）技术。该技术涉及使用核酸、植物汁液，甚至植物组织的痕迹。添加一组四个到六个引物、Bst DNA 聚合酶、AMV 或其他热稳定的逆转录酶，可以在 63°C 下孵育 20-40 分钟，无论是在开放管系统还是闭管系统中，都能快速检测到 RNA 靶标，如类病毒。
