ITS-RFLP optimization for dermatophyte identification from clinical sources in Alagoas (Brazil) versus phenotypic methods

Authors

  • Luisa Andrea Villanueva da Fonseca Microbiology Postgraduate Program, University of São Paulo, São Paulo, Brazil https://orcid.org/0000-0002-5715-636X
  • Maria Anilda Santos Araújo Tiradentes University Center, Alagoas, Brazil
  • Denise Maria Wanderlei Silva Federal University of Alagoas, Institute of Biological and Health Sciences, Alagoas, Brazil https://orcid.org/0000-0001-9641-4264
  • Fernanda Cristina de Albuquerque Maranhão Federal University of Alagoas, Institute of Biological and Health Sciences, Alagoas, Brazil https://orcid.org/0000-0002-4255-6563

DOI:

https://doi.org/10.3855/jidc.17077

Keywords:

Dermatophytes, Trichophyton, ITS-RFLP, PCR

Abstract

Introduction: Dermatophytoses are superficial mycoses, and the identification of their etiological agents is routinely performed by culture and microscopic features, which is time-consuming and relies on personnel expertise. Molecular approaches have been developed to provide faster and reliable results; therefore, this study aimed to identify dermatophytes isolated from Alagoas state patients, employing phenotypical and molecular methods.

Methodology: Clinical samples for morphological identification were collected from private and public laboratories and cultivated on Sabouraud dextrose agar. DNA extraction was followed by ITS amplicon analysis after restriction enzyme digestion DdeI (ITS-RFLP).

Results: Out of fourteen representative strains, ITS-RFLP with DdeI efficiently identified Microsporum canis, Nannizzia gypsea, and Trichophyton rubrum, while species of the complex T. tonsurans/T. mentagrophytes presented the same restriction pattern. After genotyping, 2 T. tonsurans and 1 Microsporum sp. strain were reclassified as T. rubrum.

Conclusions: RFLP of ITS-region followed by DdeI digestion produced faster and relatively reliable results than classic methods; however, this method has not been as efficient for closely related dermatophytes cryptic species.

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Author Biographies

Luisa Andrea Villanueva da Fonseca, Microbiology Postgraduate Program, University of São Paulo, São Paulo, Brazil

Master's student - Microbiology Department

Fernanda Cristina de Albuquerque Maranhão, Federal University of Alagoas, Institute of Biological and Health Sciences, Alagoas, Brazil

Professor em leadership in the Laboratory of Clinical Microbiology at UFAL.

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Published

2022-11-29

How to Cite

1.
Villanueva da Fonseca LA, Santos Araújo MA, Wanderlei Silva DM, de Albuquerque Maranhão FC (2022) ITS-RFLP optimization for dermatophyte identification from clinical sources in Alagoas (Brazil) versus phenotypic methods. J Infect Dev Ctries 16:1773–1777. doi: 10.3855/jidc.17077

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Section

Brief Original Articles