Use of restriction fragment length polymorphism to identify Candida species, related to onychomycosis
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Rasoul Mohammadi1, Parisa Badiee2, Hamid Badali3
1Department of Medical Parasitology and Mycology, Isfahan University of Medical Sciences, Isfahan, Iran.
Background:
Onychomycosis is one of the most common clinical forms of fungal infections due to both filamentous fungi and yeasts. The genus of Candida is one of the most prominent causes of onychomycosis in all around the world. Although Candida albicans is still the most frequent cause of nail infections, use of broad-spectrum antifungal agents has led to a shift in the etiology of C. albicans to non-albicans species. The aim of the present study is rapid and precise identification of candida species isolated from nail infection by using of PCR-RFLP technique.
Materials And Methods:
A total of 360 clinical yeast strains were collected from nail infections in Iran. Genomic DNA was extracted using FTA(;) cards. ITS1-5.8SrDNA-ITS2 region was amplified using universal primers and subsequently products were digested with the restriction enzyme MspI. For identification of newly described species (C. parapsilosis complex), the SADH gene was amplified, followed by digestion with Nla III restriction enzyme.
Results:
Candida albicans was the most commonly isolated species (41.1%), followed by C. parapsilosis (21.4%), C. tropicalis (12.8%), C. kefyr (9.4%), C. krusei (5.5%), C. orthopsilosis (4.1%), C. glabrata (2.8%), C. guilliermondii (1.4%), C. rugosa (0.8%), and C. lusitaniae (0.5%). Patients in the age groups of 51-60 and 81-90 years had the highest and lowest distribution of positive specimens, respectively.
Conclusion:
Rapid and precise identification of Candida species from clinical specimens lead to appropriate therapeutic plans.
Insights
This study used PCR-RFLP to rapidly identify Candida species causing onychomycosis. Candida albicans was most common, but non-albicans species are emerging, highlighting the need for precise fungal identification.
Area of Science:
- Medical Mycology
- Molecular Biology
- Clinical Microbiology
Background:
- Onychomycosis is a common fungal nail infection, frequently caused by yeasts of the genus Candida.
- Candida albicans is a primary cause, but non-albicans species are increasingly implicated in nail infections.
- Accurate identification of Candida species is crucial for effective treatment.
Purpose of the Study:
- To rapidly and precisely identify Candida species isolated from nail infections.
- To evaluate the utility of PCR-RFLP for Candida species identification.
- To understand the current etiological landscape of Candida onychomycosis.
Main Methods:
- Collected 360 clinical yeast strains from nail infections in Iran.
- Extracted genomic DNA using FTA cards.
- Amplified the ITS1-5.8SrDNA-ITS2 region and digested with MspI; amplified the SADH gene and digested with Nla III for specific species identification.
Main Results:
- Candida albicans was the most frequent isolate (41.1%).
- Non-albicans species identified included C. parapsilosis (21.4%), C. tropicalis (12.8%), and others.
- Age distribution showed the highest prevalence in the 51-60 year age group.
Conclusions:
- PCR-RFLP provides rapid and precise identification of Candida species.
- Accurate species identification is essential for guiding appropriate antifungal therapy.
- Monitoring the shift towards non-albicans species is important for public health.
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