Madurella real-time PCR, a novel approach for eumycetoma diagnosis

Amir Arastehfar (1) , Wilson Lim (2) , Farnaz Daneshnia (1) , Wendy W. J. van de Sande (2) , Ahmed H. Fahal (3) , Marie Desnos-Ollivier (4, 5) , Gerrit S. de Hoog (1, 6, 7) , Teun Boekhout (1, 8) , Sarah A. Ahmed (1, 3)

1 WI - Westerdijk Fungal Biodiversity Institute [Utrecht]
2 Erasmus MC - Erasmus University Medical Center [Rotterdam]
3 University of Khartoum
4 Mycologie moléculaire - Molecular Mycology
5 CNRMA - Centre National de Référence Mycoses Invasives et Antifongiques - National Reference Center Invasive Mycoses & Antifungals
6 Radboud University Medical Center [Nijmegen]
7 Canisius-Wilhelmina Hospital [Nijmegen, The Netherlands]
8 UvA - University of Amsterdam [Amsterdam] = Universiteit van Amsterdam

The genus Madurella comprising four species, M. fahalii, M. mycetomatis, M. pseudomycetomatis, and M. tropicana, represents the prevalent cause of eumycetoma worldwide. The four species are phenotypically similar and cause an invariable clinical picture, but differ markedly in their susceptibility to antifungal drugs, and epidemiological pattern. Therefore, specific identification is required for optimal management of Madurella infection and to reveal proper epidemiology of the species. In this study, a novel multiplex real-time PCR targeting the four Madurella species was developed and standardized. Evaluation of the assay using reference strains of the target and non-target species resulted in 100% specificity, high analytical reproducibility (R2 values >0.99) and a lowest detection limit of 3 pg target DNA. The accuracy of the real-time PCR was further assessed using biopsies from eumycetoma suspected patients. Unlike culture and DNA sequencing as gold standard diagnostic methods, the real-time PCR yielded accurate diagnosis with specific identification of the causative species in three hours compared to one or two weeks required for culture. The novel method reduces turnaround time as well as labor intensity and high costs associated with current reference methods.

Domaines: Mycologie

Amir Arastehfar, Wilson Lim, Farnaz Daneshnia, Wendy W. J. van de Sande, Ahmed H. Fahal, et al.. Madurella real-time PCR, a novel approach for eumycetoma diagnosis. PLoS Neglected Tropical Diseases, 2020, 14 (1), pp.e0007845. ⟨10.1371/journal.pntd.0007845⟩. ⟨pasteur-03243534⟩