Direct 16S/ITS rRNA Gene PCR Followed by Sanger Sequencing for Detection of Mycetoma Causative Agents in Dakar, Senegal: A Pilot Study Among Patients with Mycetoma Attending Aristide Le Dantec University Hospital.
Mycetoma
/ microbiology
Humans
RNA, Ribosomal, 16S
/ genetics
Adult
Senegal
Middle Aged
Male
Adolescent
Young Adult
Aged
Female
Polymerase Chain Reaction
/ methods
Hospitals, University
Pilot Projects
Sequence Analysis, DNA
DNA, Ribosomal Spacer
/ genetics
Madurella
/ genetics
Fungi
/ genetics
DNA, Fungal
/ genetics
Actinobacteria
/ genetics
16S
ITS
Mycetoma causative agent
PCR sequencing
Senegal
Journal
Mycopathologia
ISSN: 1573-0832
Titre abrégé: Mycopathologia
Pays: Netherlands
ID NLM: 7505689
Informations de publication
Date de publication:
09 Sep 2024
09 Sep 2024
Historique:
received:
21
05
2024
accepted:
29
08
2024
medline:
9
9
2024
pubmed:
9
9
2024
entrez:
9
9
2024
Statut:
epublish
Résumé
Mycetoma can be caused either by fungi or aerobic Actinomycetes. A precise identification of the causal agents is critical for the therapeutic outcome. Thus, this study aimed to identify the pathogens of mycetoma using 16S/ITS rRNA gene polymerase chain reaction (PCR) followed by Sanger sequencing directly on grains. In sum, 32 samples including 15 black grains, 12 red grains, and five white/yellow grains collected from patients with mycetoma at the Aristide Le Dantec University Hospital in Dakar, Senegal, between October 2014 and September 2020 were submitted to PCR/sequencing. For black grain eumycetoma, the ITS rRNA region was targeted. Similarly, the 16S rRNA gene was targeted for red grain actinomycetoma. These two regions were targeted in parallel for white/yellow grains, which could be of either bacterial or fungal origin. The age of the patients ranged from 14 to 72 years with a mean age of 36 ± 14 years. Thirteen (86%) of the 15 samples with black grains, were successfully sequenced with only one established eumycetoma pathogen, Madurella mycetomatis identified in 11 (73%). Cladosporium sphaerospermum was identified in one sample. For the 16S rRNA sequencing of red grains, a 58.3% (7/12) success rate was obtained with Actinomadura pelletieri identified in six samples. Among the five samples sequenced twice, the 16S rRNA allowed us to identify the causative agent in 2 cases, A. madurae in one, and A. geliboluensis in the other. The ITS rRNA identified 3 fungi, of which none was a mycetoma agent. Overall, direct 16S/ITS rRNA sequencing of the grains for detecting and identifying mycetoma pathogens was successful in 59.4% of cases. Fungi, led by M. mycetomatis, were the predominant pathogens identified. Two probable new mycetoma agents, C. sphaerospermum, and A. geliboluensis were identified and both deserve to be confirmed in further studies.
Identifiants
pubmed: 39249620
doi: 10.1007/s11046-024-00891-w
pii: 10.1007/s11046-024-00891-w
doi:
Substances chimiques
RNA, Ribosomal, 16S
0
DNA, Ribosomal Spacer
0
DNA, Fungal
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
80Informations de copyright
© 2024. The Author(s), under exclusive licence to Springer Nature B.V.
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