Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) at the Service of Viroid Detection.

Abstract

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 (20-40min) incubation at 63°C either in an open or close-tube system.