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<p>Root-knot nematodes (RKN - <i>Meloidogyne</i> spp.) are sedentary endoparasites that attack a wide range of plants species, including coffee. <i>Meloidogyne exigua</i>, <i>M. incognita</i>, and <i>M. paranaensis</i> are most common RKN species found in coffee-producing areas of Brazil, with <i>M. exigua</i> being the most prevalent. Effective control of these nematodes may be species-specific, necessitating accurate identification. Loop-mediated isothermal amplification (LAMP) offers a rapid, accurate, and cost-effective alternative for molecular identification of plant-parasitic nematodes. In this study, we developed a novel LAMP assay for detecting <i>M. exigua</i> by targeting the 28&#xa0;S rDNA gene. The reaction was monitored through a colorimetric change using a commercial master mix with a pH indicator and confirmed by agarose gel electrophoresis. The assay specifically detected <i>M. exigua</i> within 75&#xa0;min without cross-amplifying DNA from the other coffee root-knot nematodes, <i>M. incognita</i> and <i>M. paranaensis</i>. Additionally, the LAMP assay exhibited high sensitivity, successfully detecting 2.6&#xa0;fg µL<sup>− 1</sup> of <i>M. exigua</i> DNA. This method can be a highly effective tool for accurately identifying <i>M. exigua</i>, aiding in the diagnosis and management of this nematode.</p>

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Rapid and highly specific LAMP assay for detection of Meloidogyne exigua

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年2月9日

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  • Maria Luíza Araújo Londe,
  • Gustavo Braga Babilônia,
  • Raquel Aime Louroza Ribeiro,
  • Hevila Cristina de Brito,
  • Everaldo Antônio Lopes

摘要

Root-knot nematodes (RKN - Meloidogyne spp.) are sedentary endoparasites that attack a wide range of plants species, including coffee. Meloidogyne exigua, M. incognita, and M. paranaensis are most common RKN species found in coffee-producing areas of Brazil, with M. exigua being the most prevalent. Effective control of these nematodes may be species-specific, necessitating accurate identification. Loop-mediated isothermal amplification (LAMP) offers a rapid, accurate, and cost-effective alternative for molecular identification of plant-parasitic nematodes. In this study, we developed a novel LAMP assay for detecting M. exigua by targeting the 28 S rDNA gene. The reaction was monitored through a colorimetric change using a commercial master mix with a pH indicator and confirmed by agarose gel electrophoresis. The assay specifically detected M. exigua within 75 min without cross-amplifying DNA from the other coffee root-knot nematodes, M. incognita and M. paranaensis. Additionally, the LAMP assay exhibited high sensitivity, successfully detecting 2.6 fg µL− 1 of M. exigua DNA. This method can be a highly effective tool for accurately identifying M. exigua, aiding in the diagnosis and management of this nematode.