Specific and sensitive loop-mediated isothermal amplification (LAMP) method for Madurella strains, eumycetoma filamentous fungi causative agent

Isato Yoshioka, Yugo Mori, Ahmed Hassan Fahal, Emmanuel Edwar Siddig, Satoshi Kaneko, Takashi Yaguchi*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

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Abstract

Background Filamentous fungi of the genus Madurella are the primary causative agents of mycetoma, a disease observed in tropical and subtropical regions. Since early diagnostics based on a morphological approach are difficult and have many shortcomings, a molecular diagnostic method suitable for rural settings is required. In this study, we developed the loop-mediated isothermal amplification (LAMP) method to present a foundational technique of the diagnosis of Madurella spp. (M. mycetomatis, M. pseudomycetomatis, M. tropicana, and M. faha-lii), the common causative organisms of eumycetoma. Principal findings We successfully designed a primer pair targeting the rDNAs of three Madurella spp. exclud-ing M. fahalii, and detected up to 100 fg of genomic DNA extracted from isolates of M. myce-tomatis and 1 pg of M. pseudomycetomatis and M. tropicana, within one hour. Second, a primer pair specific to M. mycetomatis, the most common causative species, or M. fahalii, a drug-resistant species, was constructed, and the detection limit of both primer pairs was 1 pg. The designed primers accurately distinguished 16 strains of the genus Madurella from various fungal species known to cause mycetomas. Conclusion In summary, we established the first model of a LAMP detection method that rapidly and sensitively detects and identifies Madurella isolates for clinical diagnostics. Moreover, the combined designed primer sets could identify mycetoma-causing strains simultaneously.

Original languageEnglish
Article numbere0011644
JournalPLoS Neglected Tropical Diseases
Volume17
Issue number9 September
DOIs
Publication statusPublished - 18 Sept 2023

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Publisher Copyright: © 2023 Yoshioka et al.

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