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

Amir Arastehfar, Wilson Lim, Farnaz Daneshnia, Wendy W J van de Sande, Ahmed H Fahal, Marie Desnos-Ollivier, Gerrit S de Hoog, Teun Boekhout, Sarah A Ahmed

Onderzoeksoutput: Bijdrage aan wetenschappelijk tijdschrift/periodieke uitgaveArtikelWetenschappelijkpeer review

Samenvatting

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.

Originele taal-2Engels
Pagina's (van-tot)e0007845
TijdschriftPLoS Neglected Tropical Diseases
Volume14
Nummer van het tijdschrift1
DOI's
StatusGepubliceerd - 15 jan 2020

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    Arastehfar, A., Lim, W., Daneshnia, F., van de Sande, W. W. J., Fahal, A. H., Desnos-Ollivier, M., de Hoog, G. S., Boekhout, T., & Ahmed, S. A. (2020). Madurella real-time PCR, a novel approach for eumycetoma diagnosis. PLoS Neglected Tropical Diseases, 14(1), e0007845. https://doi.org/10.1371/journal.pntd.0007845