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<p>Succinate dehydrogenase inhibitors (SDHIs) are widely used to control <i>Botrytis cinerea</i>, but resistance caused by multiple point mutations in the <i>sdh</i> genes has become a major concern. To enable rapid and accurate identification of SDHI‑resistant genotypes, we developed a loop‑mediated isothermal amplification (LAMP) assay using a universal quenching probe (UQProbe LAMP) capable of simultaneously detecting multiple single nucleotide polymorphisms (SNPs) within the same target region. Using a single UQProbe and a common LAMP primer set, the method successfully distinguished four <i>sdh</i>B genotypes—wild type, H272R, H272Y, and H272V—based on characteristic melting temperatures (approximately 56&#xa0;°C, 50&#xa0;°C, 44.5&#xa0;°C, and 38.5&#xa0;°C, respectively). This single‑probe, multi‑SNP detection strategy eliminates the need for mutation‑specific primers or multiple probes, providing a simple and efficient workflow for SNP typing. The UQProbe LAMP system developed in this study offers a rapid, cost‑effective, and versatile platform for multiplex SNP discrimination, applicable not only to fungicide resistance monitoring but also to a wide range of genetic diagnostic applications requiring precise SNP detection.</p> Graphical abstract <p></p>

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Simultaneous detection of multiple sdhB SNPs in Botrytis cinerea using a universal quenching probe–based LAMP assay

  • 期刊论文

  • 发表日期：

    2026年6月15日

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  • Shiro Fukuta,
  • Hiroko Kojima-Ando,
  • Ken-ichiro Mori,
  • Tomoaki Hara,
  • Masami Tsuzuki,
  • Reina Inagaki,
  • Kenta Tsunekawa,
  • Yuta Uchida,
  • Mizuho Arakawa,
  • Kensuke Yamada

摘要

Succinate dehydrogenase inhibitors (SDHIs) are widely used to control Botrytis cinerea, but resistance caused by multiple point mutations in the sdh genes has become a major concern. To enable rapid and accurate identification of SDHI‑resistant genotypes, we developed a loop‑mediated isothermal amplification (LAMP) assay using a universal quenching probe (UQProbe LAMP) capable of simultaneously detecting multiple single nucleotide polymorphisms (SNPs) within the same target region. Using a single UQProbe and a common LAMP primer set, the method successfully distinguished four sdhB genotypes—wild type, H272R, H272Y, and H272V—based on characteristic melting temperatures (approximately 56 °C, 50 °C, 44.5 °C, and 38.5 °C, respectively). This single‑probe, multi‑SNP detection strategy eliminates the need for mutation‑specific primers or multiple probes, providing a simple and efficient workflow for SNP typing. The UQProbe LAMP system developed in this study offers a rapid, cost‑effective, and versatile platform for multiplex SNP discrimination, applicable not only to fungicide resistance monitoring but also to a wide range of genetic diagnostic applications requiring precise SNP detection.

Graphical abstract