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Use of Restriction Fragment Length Polymorphism to Rapidly Identify Dermatophyte Species Related to Dermatophytosis

BACKGROUND: Dermatophytes are a group of keratinophilic fungi worldwide, which can infect the skin, hair and nails of humans and animals. This genus includes several species that present different features of dermatophytosis. Although, laboratory diagnosis of dermatophytes is based on direct microsc...

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Autores principales: Mohammadi, Rasoul, Abastabar, Mahdi, Mirhendi, Hossein, Badali, Hamid, Shadzi, Shahla, Chadeganipour, Mustafa, Pourfathi, Parinaz, Jalalizand, Niloufar, Haghani, Iman
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Kowsar 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4541063/
https://www.ncbi.nlm.nih.gov/pubmed/26301058
http://dx.doi.org/10.5812/jjm.8(5)2015.17296
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author Mohammadi, Rasoul
Abastabar, Mahdi
Mirhendi, Hossein
Badali, Hamid
Shadzi, Shahla
Chadeganipour, Mustafa
Pourfathi, Parinaz
Jalalizand, Niloufar
Haghani, Iman
author_facet Mohammadi, Rasoul
Abastabar, Mahdi
Mirhendi, Hossein
Badali, Hamid
Shadzi, Shahla
Chadeganipour, Mustafa
Pourfathi, Parinaz
Jalalizand, Niloufar
Haghani, Iman
author_sort Mohammadi, Rasoul
collection PubMed
description BACKGROUND: Dermatophytes are a group of keratinophilic fungi worldwide, which can infect the skin, hair and nails of humans and animals. This genus includes several species that present different features of dermatophytosis. Although, laboratory diagnosis of dermatophytes is based on direct microscopy, biochemical tests and culture, these manners are expensive, time consuming and need skilled staff. Therefore, molecular methods like PCR-RFLP are the beneficial tools for identification, which are rapid and sensitive. Thus, dermatophyte species are able to generate characteristic band patterns on agarose gel electrophoresis using PCR-RFLP technique, which leads to successful identification at the species level within a 5-hour period. OBJECTIVES: The purpose of this study was to study inter- and intraspecific genomic variations for identification of clinically important dermatophyte species obtained from clinical specimens in Isfahan, Iran using PCR-RFLP. MATERIALS AND METHODS: From March 2011 to August 2012, 135 clinical isolates were collected from infected patients at Isfahan, Iran. ITS1-5.8S-ITS2 region of rDNA was amplified using universal fungal primers. Subsequently, amplified products were digested by the MvaI restriction enzyme. Using discriminating band profiles on agarose gel, dermatophyte species were identified. However, DNA sequencing was used for unidentifiable strains. RESULTS: The specimens were obtained from skin scrapings (70.3%), nail (24.4%) and hair (5.1%) clippings. Most patients were between 21 - 30 years and the ratio of male to female was 93/42. Trichophyton interdigitale was the commonest isolate (52.5%) in our findings, followed by Epidermophyton floccosum (24.4%), T. rubrum (16.2%), Microsporum canis (2.2%), T. erinacei (1.4%), T. violaceum (1.4%), T. tonsurans (0.7%) and M. gypseum (0.7%) based on PCR-RFLP. CONCLUSIONS: Combination of traditional methods and molecular techniques considerably improves identification of dermatophytes in the species level in clinical laboratories, which can lead to properly antifungal therapy and successful management of infections. However, restriction and specificity and sensitivity should be lowered and increased, respectively, to be useful for a wide variety of clinical applications.
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spelling pubmed-45410632015-08-21 Use of Restriction Fragment Length Polymorphism to Rapidly Identify Dermatophyte Species Related to Dermatophytosis Mohammadi, Rasoul Abastabar, Mahdi Mirhendi, Hossein Badali, Hamid Shadzi, Shahla Chadeganipour, Mustafa Pourfathi, Parinaz Jalalizand, Niloufar Haghani, Iman Jundishapur J Microbiol Research Article BACKGROUND: Dermatophytes are a group of keratinophilic fungi worldwide, which can infect the skin, hair and nails of humans and animals. This genus includes several species that present different features of dermatophytosis. Although, laboratory diagnosis of dermatophytes is based on direct microscopy, biochemical tests and culture, these manners are expensive, time consuming and need skilled staff. Therefore, molecular methods like PCR-RFLP are the beneficial tools for identification, which are rapid and sensitive. Thus, dermatophyte species are able to generate characteristic band patterns on agarose gel electrophoresis using PCR-RFLP technique, which leads to successful identification at the species level within a 5-hour period. OBJECTIVES: The purpose of this study was to study inter- and intraspecific genomic variations for identification of clinically important dermatophyte species obtained from clinical specimens in Isfahan, Iran using PCR-RFLP. MATERIALS AND METHODS: From March 2011 to August 2012, 135 clinical isolates were collected from infected patients at Isfahan, Iran. ITS1-5.8S-ITS2 region of rDNA was amplified using universal fungal primers. Subsequently, amplified products were digested by the MvaI restriction enzyme. Using discriminating band profiles on agarose gel, dermatophyte species were identified. However, DNA sequencing was used for unidentifiable strains. RESULTS: The specimens were obtained from skin scrapings (70.3%), nail (24.4%) and hair (5.1%) clippings. Most patients were between 21 - 30 years and the ratio of male to female was 93/42. Trichophyton interdigitale was the commonest isolate (52.5%) in our findings, followed by Epidermophyton floccosum (24.4%), T. rubrum (16.2%), Microsporum canis (2.2%), T. erinacei (1.4%), T. violaceum (1.4%), T. tonsurans (0.7%) and M. gypseum (0.7%) based on PCR-RFLP. CONCLUSIONS: Combination of traditional methods and molecular techniques considerably improves identification of dermatophytes in the species level in clinical laboratories, which can lead to properly antifungal therapy and successful management of infections. However, restriction and specificity and sensitivity should be lowered and increased, respectively, to be useful for a wide variety of clinical applications. Kowsar 2015-06-30 /pmc/articles/PMC4541063/ /pubmed/26301058 http://dx.doi.org/10.5812/jjm.8(5)2015.17296 Text en Copyright © 2015, Ahvaz Jundishapur University of Medical Sciences. http://creativecommons.org/licenses/by-nc/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License (http://creativecommons.org/licenses/by-nc/4.0/) which permits copy and redistribute the material just in noncommercial usages, provided the original work is properly cited.
spellingShingle Research Article
Mohammadi, Rasoul
Abastabar, Mahdi
Mirhendi, Hossein
Badali, Hamid
Shadzi, Shahla
Chadeganipour, Mustafa
Pourfathi, Parinaz
Jalalizand, Niloufar
Haghani, Iman
Use of Restriction Fragment Length Polymorphism to Rapidly Identify Dermatophyte Species Related to Dermatophytosis
title Use of Restriction Fragment Length Polymorphism to Rapidly Identify Dermatophyte Species Related to Dermatophytosis
title_full Use of Restriction Fragment Length Polymorphism to Rapidly Identify Dermatophyte Species Related to Dermatophytosis
title_fullStr Use of Restriction Fragment Length Polymorphism to Rapidly Identify Dermatophyte Species Related to Dermatophytosis
title_full_unstemmed Use of Restriction Fragment Length Polymorphism to Rapidly Identify Dermatophyte Species Related to Dermatophytosis
title_short Use of Restriction Fragment Length Polymorphism to Rapidly Identify Dermatophyte Species Related to Dermatophytosis
title_sort use of restriction fragment length polymorphism to rapidly identify dermatophyte species related to dermatophytosis
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4541063/
https://www.ncbi.nlm.nih.gov/pubmed/26301058
http://dx.doi.org/10.5812/jjm.8(5)2015.17296
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