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Madurella real-time PCR, a novel approach for eumycetoma diagnosis
- Source :
- 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)
- Publication Year :
- 2020
- Publisher :
- Public Library of Science, 2020.
-
Abstract
- 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.<br />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.
- Subjects :
- 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
Subjects
Details
- Language :
- English
- ISSN :
- 19352735 and 19352727
- Volume :
- 14
- Database :
- OpenAIRE
- Journal :
- PLoS Neglected Tropical Diseases
- Accession number :
- edsair.doi.dedup.....2e47fd7b9af299a748b6152fd988cd2e
- Full Text :
- https://doi.org/10.1371/journal.pntd.0007845