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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.

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

  • 实验方案

  • 发表日期：

    2026年

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

摘要

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.