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Whole Genome Sequencing of Candida glabrata for Detection of Markers of Antifungal Drug Resistance
Published on: December 28, 2017
The new mutation L321F in Candida albicans ERG11 gene may be associated with fluconazole resistance
Vagner Oliveira Carvalho1, Thelma S Okay, Márcia S C Melhem
1Laboratory of Medical Mycology (LIM-53)-Clinical Dermatology Division, Hospital das Clínicas da FMUSP e Instituto de Medicina Tropical de São Paulo, Universidade de São Paulo, São Paulo, State of São Paulo, Brazil.
Background:
For many years fluconazole has been commonly used to treat Candida infections. However, the indiscriminate use of this antimycotic therapy has favored the emergence of resistant isolates. Mutations in the ERG11 gene have been described as one of the primary mechanisms of resistance in Candida species.
Aims:
In this study we investigated missense mutations in ERG11 genes of Candida albicans, Candida glabrata and Candida tropicalis isolates previously evaluated by susceptibility testing to fluconazole.
Methods:
Screening for these mutations was performed on 19 Candida clinical isolates (eight C. albicans, five C. glabrata and six C. tropicalis) resistant and susceptible to fluconazole. The ERG11 gene was amplified by PCR with specific primers for each Candida species and analyzed by automated sequencing.
Results:
We identified 14 different missense mutations, five of which had not been described previously. Among them, a new mutation L321F was identified in a fluconazole resistant C. albicans isolate and it was analyzed by a theoretical three-dimensional structure of the ERG11p.
Conclusion:
The L321F mutation in C. albicans ERG11 gene may be associated with fluconazole resistance.
Insights
New mutations in the ERG11 gene are emerging in Candida species, contributing to fluconazole resistance. A novel L321F mutation in Candida albicans may be linked to this antifungal resistance.
Area of Science:
- Mycology
- Molecular Biology
- Antimicrobial Resistance
Background:
- Fluconazole is a widely used antifungal for Candida infections.
- Increased use has led to the emergence of fluconazole-resistant Candida isolates.
- Mutations in the ERG11 gene are a key mechanism for this resistance.
Purpose of the Study:
- To investigate missense mutations in the ERG11 gene of fluconazole-resistant and susceptible Candida albicans, Candida glabrata, and Candida tropicalis.
- To identify novel mutations associated with antifungal resistance.
Main Methods:
- Analysis of 19 clinical Candida isolates (8 C. albicans, 5 C. glabrata, 6 C. tropicalis) with known fluconazole susceptibility.
- PCR amplification of the ERG11 gene using species-specific primers.
- Automated sequencing of the ERG11 gene to identify missense mutations.
Main Results:
- Identified 14 distinct missense mutations in the ERG11 gene.
- Five of these mutations were previously undescribed.
- A novel mutation, L321F, was found in a fluconazole-resistant Candida albicans isolate.
- The L321F mutation was further analyzed using a theoretical 3D structure of the ERG11 protein.
Conclusions:
- The novel L321F mutation in the ERG11 gene of Candida albicans is potentially associated with fluconazole resistance.
- This finding contributes to understanding the molecular mechanisms of antifungal resistance in Candida species.
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