Madurella real-time PCR, a novel approach for eumycetoma diagnosis.
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 t...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2020-01-01
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| Series: | PLoS Neglected Tropical Diseases |
| Online Access: | https://doi.org/10.1371/journal.pntd.0007845 |
| _version_ | 1819112918495526912 |
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| author | 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 |
| author_facet | 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 |
| author_sort | Amir Arastehfar |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-22T04:21:08Z |
| format | Article |
| id | doaj.art-1e649db94212483d9dc32a3e88c58780 |
| institution | Directory Open Access Journal |
| issn | 1935-2727 1935-2735 |
| language | English |
| last_indexed | 2024-12-22T04:21:08Z |
| publishDate | 2020-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Neglected Tropical Diseases |
| spelling | doaj.art-1e649db94212483d9dc32a3e88c587802022-12-21T18:39:17ZengPublic Library of Science (PLoS)PLoS Neglected Tropical Diseases1935-27271935-27352020-01-01141e000784510.1371/journal.pntd.0007845Madurella real-time PCR, a novel approach for eumycetoma diagnosis.Amir ArastehfarWilson LimFarnaz DaneshniaWendy W J van de SandeAhmed H FahalMarie Desnos-OllivierGerrit S de HoogTeun BoekhoutSarah A AhmedThe 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.https://doi.org/10.1371/journal.pntd.0007845 |
| spellingShingle | 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 Madurella real-time PCR, a novel approach for eumycetoma diagnosis. PLoS Neglected Tropical Diseases |
| title | Madurella real-time PCR, a novel approach for eumycetoma diagnosis. |
| title_full | Madurella real-time PCR, a novel approach for eumycetoma diagnosis. |
| title_fullStr | Madurella real-time PCR, a novel approach for eumycetoma diagnosis. |
| title_full_unstemmed | Madurella real-time PCR, a novel approach for eumycetoma diagnosis. |
| title_short | Madurella real-time PCR, a novel approach for eumycetoma diagnosis. |
| title_sort | madurella real time pcr a novel approach for eumycetoma diagnosis |
| url | https://doi.org/10.1371/journal.pntd.0007845 |
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