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Molecular characterization of Alternaria alternata causing ocular infection: detection of IGS-RFLP intraspecific
Federico S Laich1, Julia Alcoba-Flórez, Eduardo Pérez-Roth
1Unidad de Investigación, Asociada al Centro de Investigaciones Biológicas, CSIC, Madrid, Spain.
Abstract:
A case of fungal keratitis is presented in which corneal scrapings were obtained for microbiological studies, including morphological identification and molecular characterization of the etiologic agent. Comparative sequence analyses of the Internal Transcribed Spacer domain of 5.8S and 26S regions of nuclear rDNA showed 100% identity with different species of Alternaria and PCR-RFLP analysis of Intergenic Spacer regions revealed intraspecific variation. The combination of morphological and molecular characters resulted in the unambiguous identification of the causal agent as Alternaria alternata. Treatment with antifungals contributed to the improvement in the patient's lesions.
Insights
This study identified Alternaria alternata as the cause of fungal keratitis using molecular and morphological methods. Antifungal treatment effectively improved the patient's corneal lesions.
Area of Science:
- Ophthalmology
- Mycology
- Molecular Biology
Background:
- Fungal keratitis is a serious eye infection that can lead to vision loss.
- Accurate identification of the causative fungal species is crucial for effective treatment.
Observation:
- A patient presented with fungal keratitis, necessitating microbiological investigation of corneal scrapings.
- Morphological and molecular techniques were employed to identify the fungal pathogen.
Findings:
- Comparative sequence analysis of nuclear ribosomal DNA (rDNA) Internal Transcribed Spacer (ITS) regions showed 100% identity with various Alternaria species.
- Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) analysis of Intergenic Spacer (IGS) regions revealed intraspecific variation within the Alternaria isolates.
- The combined morphological and molecular data unambiguously identified the etiological agent as Alternaria alternata.
Implications:
- This case highlights the importance of integrated diagnostic approaches for accurate fungal keratitis diagnosis.
- Successful treatment with antifungals underscores the efficacy of targeted therapy once the pathogen is identified.
- The findings contribute to understanding the molecular diversity of Alternaria species causing ocular infections.
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