ITS-RFLP optimization for dermatophyte identification from clinical sources in Alagoas (Brazil) versus phenotypic methods
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 re...
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The Journal of Infection in Developing Countries
2022-11-01
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| Series: | Journal of Infection in Developing Countries |
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| Online Access: | https://jidc.org/index.php/journal/article/view/17077 |
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| author | Luisa Andrea Villanueva da Fonseca Maria Anilda Santos Araújo Denise Maria Wanderlei Silva Fernanda Cristina de Albuquerque Maranhão |
| author_facet | Luisa Andrea Villanueva da Fonseca Maria Anilda Santos Araújo Denise Maria Wanderlei Silva Fernanda Cristina de Albuquerque Maranhão |
| author_sort | Luisa Andrea Villanueva da Fonseca |
| collection | DOAJ |
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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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| format | Article |
| id | doaj-art-8eff2c93e9104c6a9994cb9c29ff8dab |
| institution | DOAJ |
| issn | 1972-2680 |
| language | English |
| publishDate | 2022-11-01 |
| publisher | The Journal of Infection in Developing Countries |
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| series | Journal of Infection in Developing Countries |
| spelling | doaj-art-8eff2c93e9104c6a9994cb9c29ff8dab2025-08-20T02:57:52ZengThe Journal of Infection in Developing CountriesJournal of Infection in Developing Countries1972-26802022-11-01161110.3855/jidc.17077ITS-RFLP optimization for dermatophyte identification from clinical sources in Alagoas (Brazil) versus phenotypic methodsLuisa Andrea Villanueva da Fonseca0Maria Anilda Santos Araújo1Denise Maria Wanderlei Silva2Fernanda Cristina de Albuquerque Maranhão3Microbiology Postgraduate Program, University of São Paulo, São Paulo, BrazilTiradentes University Center, Alagoas, BrazilFederal University of Alagoas, Institute of Biological and Health Sciences, Alagoas, BrazilFederal University of Alagoas, Institute of Biological and Health Sciences, Alagoas, Brazil 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. https://jidc.org/index.php/journal/article/view/17077DermatophytesTrichophytonITS-RFLPPCR |
| spellingShingle | Luisa Andrea Villanueva da Fonseca Maria Anilda Santos Araújo Denise Maria Wanderlei Silva Fernanda Cristina de Albuquerque Maranhão ITS-RFLP optimization for dermatophyte identification from clinical sources in Alagoas (Brazil) versus phenotypic methods Journal of Infection in Developing Countries Dermatophytes Trichophyton ITS-RFLP PCR |
| title | ITS-RFLP optimization for dermatophyte identification from clinical sources in Alagoas (Brazil) versus phenotypic methods |
| title_full | ITS-RFLP optimization for dermatophyte identification from clinical sources in Alagoas (Brazil) versus phenotypic methods |
| title_fullStr | ITS-RFLP optimization for dermatophyte identification from clinical sources in Alagoas (Brazil) versus phenotypic methods |
| title_full_unstemmed | ITS-RFLP optimization for dermatophyte identification from clinical sources in Alagoas (Brazil) versus phenotypic methods |
| title_short | ITS-RFLP optimization for dermatophyte identification from clinical sources in Alagoas (Brazil) versus phenotypic methods |
| title_sort | its rflp optimization for dermatophyte identification from clinical sources in alagoas brazil versus phenotypic methods |
| topic | Dermatophytes Trichophyton ITS-RFLP PCR |
| url | https://jidc.org/index.php/journal/article/view/17077 |
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