Madurella real-time PCR, a novel approach for eumycetoma diagnosis
| Autor: | Wendy W. J. van de Sande, Wilson Lim, Gerrit S. de Hoog, Ahmed H. Fahal, Sarah A. Ahmed, Farnaz Daneshnia, Amir Arastehfar, Marie Desnos-Ollivier, Teun Boekhout |
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| Přispěvatelé: | Westerdijk Fungal Biodiversity Institute [Utrecht] (WI), Royal Netherlands Academy of Arts and Sciences (KNAW), Erasmus University Medical Center [Rotterdam] (Erasmus MC), University of Khartoum, Mycologie moléculaire - Molecular Mycology, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Centre National de Référence Mycoses Invasives et Antifongiques - National Reference Center Invasive Mycoses & Antifungals (CNRMA), Institut Pasteur [Paris] (IP), Radboud University Medical Center [Nijmegen], Canisius-Wilhelmina Hospital [Nijmegen, The Netherlands], University of Amsterdam [Amsterdam] (UvA), Amir Arastehfar received grants from European Union’s Horizon 2020 research and innovation program under the Marie Sklodowska-Curie grant agreement No 642095., European Project: 642095,H2020,H2020-MSCA-ITN-2014,OPATHY(2015), Westerdijk Fungal Biodiversity Institute, Evolutionary and Population Biology (IBED, FNWI), Westerdijk Fungal Biodiversity Insitute [Utrecht] (WI), Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], Institut Pasteur [Paris], Radboud University Medical Centre [Nijmegen, The Netherlands], Medical Microbiology & Infectious Diseases |
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Madurella Biopsy RC955-962 Invasive Species Artificial Gene Amplification and Extension Pathology and Laboratory Medicine Polymerase Chain Reaction law.invention Geographical Locations Sudan MESH: Biopsy 0302 clinical medicine law Arctic medicine. Tropical medicine Medicine and Health Sciences Multiplex DNA Fungal Polymerase chain reaction [SDV.MP.MYC]Life Sciences [q-bio]/Microbiology and Parasitology/Mycology Fungal Pathogens biology Antimicrobials MESH: Real-Time Polymerase Chain Reaction Fungal Diseases Drugs 3. Good health MESH: Mycetoma Infectious Diseases Real-time polymerase chain reaction Medical Microbiology Public aspects of medicine RA1-1270 Pathogens Research Article Neglected Tropical Diseases 030231 tropical medicine Surgical and Invasive Medical Procedures Mycology Eumycetoma Real-Time Polymerase Chain Reaction Research and Analysis Methods Sensitivity and Specificity Microbiology DNA sequencing 03 medical and health sciences Species Colonization Diagnostic Medicine Microbial Control medicine MESH: Madurella Humans Molecular Biology Techniques Microbial Pathogens Molecular Biology Pharmacology Antifungals MESH: Humans Ecology and Environmental Sciences Public Health Environmental and Occupational Health Biology and Life Sciences Gold standard (test) Tropical Diseases medicine.disease biology.organism_classification Virology MESH: Sensitivity and Specificity MESH: DNA Fungal 030104 developmental biology Mycetoma People and Places Africa Specific identification |
| Zdroj: | PLoS Neglected Tropical Diseases PLoS Neglected Tropical Diseases, 2020, 14 (1), pp.e0007845. ⟨10.1371/journal.pntd.0007845⟩ PLoS Neglected Tropical Diseases, 14(1). Public Library of Science PLoS Neglected Tropical Diseases, 14(1):e0007845. Public Library of Science PLoS Neglected Tropical Diseases, Public Library of Science, 2020, 14 (1), pp.e0007845. ⟨10.1371/journal.pntd.0007845⟩ PLoS Neglected Tropical Diseases, 14. Public Library of Science PLoS Neglected Tropical Diseases, Vol 14, Iss 1, p e0007845 (2020) |
| ISSN: | 1935-2735 1935-2727 |
| DOI: | 10.1371/journal.pntd.0007845 |
| Popis: | 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. Author summary Mycetoma, a progressive and disfiguring disease, is one of the neglected tropical diseases, caused by both bacteria and fungi. Eumycetoma is the fungal type and mainly caused by species of the genus Madurella. Madurella mycetomatis is the most prevalent species worldwide. However, other species such as M. fahalii, M. pseudomycetomatis, and M. tropicana can also cause mycetoma and have a different susceptibility towards the drug used for treating mycetoma patients. Currently, we lack a rapid and non-culture-based technique that can readily identify these four species from clinical samples. Due to its sensitivity, and specificity, real-time PCR is re-recognized by European Organization for Research and Treatment of Cancer (EORTC) to directly identify fungal agents from clinical samples. We developed and validated a multiplex real-time PCR-based technique using the least expensive chemistry to identify Madurella species within 3–4 hours. Development of such a technique will allow rapid diagnosis of eumycetoma and timely initiation of appropriate antifungal therapy. |
| Databáze: | OpenAIRE |
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