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Updated: Jul 6, 2026
On-Site Molecular Detection of Soil-Borne Phytopathogens Using a Portable Real-Time PCR System
Published on: February 23, 2018
On-Site Universal Phytoplasma Detection by 23S rRNA-Gene-Targeted Loop-Mediated Isothermal Amplification (LAMP)
Mako Akahori1, Akio Miyazaki1, Hiroaki Koinuma2
1Department of Agricultural and Environmental Biology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, Japan.
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This chapter describes a genus-universal, specific, sensitive, rapid, and simple detection system for phytoplasmas using universal loop-mediated isothermal amplification (LAMP) primers targeting the 23S rRNA gene. Diagnosis with this system consists of a simple boiling DNA extraction method (95 °C, 10 min) and DNA amplification by the LAMP assay (64 °C, 30-60 min), with a total time of about 60 min. This system enables on-site diagnosis, as it proceeds at a constant temperature, is easy to operate, requires no experimental devices or electricity, and generates little experimental waste. Moreover, the simple boiling DNA extraction method can be applied to a variety of sample types, including herbaceous plants, woody plants, and insect vectors.
