On-Site Universal Phytoplasma Detection by 23S rRNA-Gene-Targeted Loop-Mediated Isothermal Amplification (LAMP).

Mako Akahori, Akio Miyazaki, Hiroaki Koinuma, Nozomu Iwabuchi, Kensaku Maejima, Shigetou Namba, Yasuyuki Yamaji

Journal: Methods in molecular biology (Clifton, N.J.) 2026;3008():75-87

PMID: 41528701

Abstract

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.

Address: Department of Agricultural and Environmental Biology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, Japan.; Department of Agriculture, School of Agriculture, Tokai University, Kumamoto, Japan.; Department of Agricultural and Environmental Biology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, Japan. [email protected].

DOI: 10.1007/978-1-0716-5104-9_7

MeSH Terms: Phytoplasma, Nucleic Acid Amplification Techniques, RNA, Ribosomal, 23S, Molecular Diagnostic Techniques, DNA, Bacterial, Plant Diseases, DNA Primers