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On-Site Molecular Detection of Soil-Borne Phytopathogens Using a Portable Real-Time PCR System
Published on: February 23, 2018
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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.
Methods in Molecular Biology (Clifton, N.J.)
|January 13, 2026
Summary
A new phytoplasma detection system uses universal loop-mediated isothermal amplification (LAMP) primers for rapid, on-site diagnosis. This simple method, combining boiling DNA extraction and LAMP assay, works for various plant and insect samples.
Area of Science:
- Plant Pathology
- Molecular Biology
- Microbial Diagnostics
Background:
- Phytoplasmas are plant pathogens causing significant agricultural losses.
- Accurate and rapid detection of phytoplasmas is crucial for disease management.
- Existing detection methods can be time-consuming, require specialized equipment, or lack broad applicability.
Purpose of the Study:
- To develop a genus-universal, specific, sensitive, rapid, and simple detection system for phytoplasmas.
- To enable on-site diagnosis of phytoplasma infections in various host types.
- To provide an accessible diagnostic tool with minimal resource requirements.
Main Methods:
- Utilized universal loop-mediated isothermal amplification (LAMP) primers targeting the 23S rRNA gene for phytoplasma detection.
- Implemented a simple boiling DNA extraction method (95°C, 10 min) suitable for diverse sample types.
- Performed DNA amplification using the LAMP assay at a constant temperature (64°C, 30-60 min).
Main Results:
- The developed system demonstrated genus-universal specificity, sensitivity, and rapidity for phytoplasma detection.
- The entire diagnostic process, including DNA extraction and amplification, takes approximately 60 minutes.
- The simple boiling DNA extraction method proved effective for herbaceous plants, woody plants, and insect vectors.
Conclusions:
- The described phytoplasma detection system offers a rapid, simple, and sensitive diagnostic solution.
- Its isothermal nature and minimal equipment requirements facilitate on-site diagnostics.
- The system's versatility across different sample types enhances its practical application in plant disease management.
