Simultaneous Detection of Four Madurella Species Using Loop-Mediated Isothermal Amplification (LAMP) for Eumycetoma Diagnosis

Eumycetoma is a neglected tropical disease caused primarily by a Madurella mycetomatis infection, besides other related species. In this study, we designed a novel loop-mediated isothermal amplification (LAMP) primer set capable of simultaneously detecting four Madurella species (M. mycetomatis, M. pseudomycetomatis, M. tropicana, and M. fahalii). Genomic sequencing of M. pseudomycetomatis strain and comparative genome analysis revealed the candidate genes that were common among and specific to Madurella species. The 3 LAMP primer sets targeting these genes detected up to 1 pg of the genomic DNA of all 4 Madurella species, exhibiting no cross-reactivity toward other pathogenic fungi. Among these, one primer set showing better reactivity was selected as a candidate used for diagnosis. Therefore, we developed novel primer sets which enabled the simultaneous detection of four Madurella species. Our present findings will lead to a faster and simpler diagnostic tool for eumycetoma detection, especially in rural clinical settings.