This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Shehata, A. S.
Right arrow Articles by Ghannoum, M. A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Shehata, A. S.
Right arrow Articles by Ghannoum, M. A.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, August 2008, p. 2641-2645, Vol. 46, No. 8
0095-1137/08/$08.00+0     doi:10.1128/JCM.00697-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Single-Step PCR Using (GACA)4 Primer: Utility for Rapid Identification of Dermatophyte Species and Strains{triangledown}

Atef S. Shehata,1,3 Pranab K. Mukherjee,3 Hassan N. Aboulatta,1 Atef I. El Akhras,2 Said H. Abbadi,1 and Mahmoud A. Ghannoum3*

Microbiology Department, Faculty of Medicine, Suez Canal University, Ismailia, Egypt,1 Dermatology Department, Faculty of Medicine, Suez Canal University, Ismailia, Egypt,2 Center for Medical Mycology, Department of Dermatology, University Hospitals of Cleveland, and Case Western Reserve University, Cleveland, Ohio3

Received 11 April 2008/ Returned for modification 7 June 2008/ Accepted 18 June 2008

Dermatophytes are fungi that belong to three genera: Epidermophyton, Microsporum, and Trichophyton. Identification of dermatophyte species is essential for appropriate diagnosis and treatment of dermatophytosis. Routine identification depends on macroscopic and microscopic morphology, which is time-consuming and does not identify dermatophyte strains. In this study, two PCR-based methods were compared for their abilities to identify 21 dermatophyte isolates obtained from Egyptian patients to the species and strain levels. The first method employed a two-step method: PCR amplification, using ITS1 and ITS4 as primers, followed by restriction enzyme digestion using the endonuclease MvaI. The second method employed a one-step approach employing the repetitive oligonucleotide (GACA)4 as a primer. Dermatophyte strains were also identified using a conventional culture method. Our results showed that the conventional culture method identified four species: Microsporum canis, Trichophyton mentagrophytes, Trichophyton rubrum, and Trichophyton violaceum. Moreover, both PCR methods agreed with the diagnosis made using the conventional approach. Furthermore, ITS1/ITS4-based PCR provided no strain differentiation, while (GACA)4-based PCR identified different varieties among the T. mentagrophytes isolates. Taken together, our results suggest that (GACA)4-based PCR has utility as a simple and rapid method for identification of dermatophyte species as well as utility for differentiation of T. mentagrophytes variants.

* Corresponding author. Mailing address: Center for Medical Mycology, Department of Dermatology, University Hospitals of Cleveland, Case Western Reserve University, 11100 Euclid Avenue, Cleveland, OH 44106. Phone: (216) 844-8580. Fax: (216) 844-1076. E-mail: Mahmoud.Ghannoum{at}case.edu

{triangledown} Published ahead of print on 25 June 2008.

Journal of Clinical Microbiology, August 2008, p. 2641-2645, Vol. 46, No. 8
0095-1137/08/$08.00+0     doi:10.1128/JCM.00697-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»