Molecular identification of dermatophytes by arbitrarily primed polymerase chain reaction (original) (raw)
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Application of PCR to Distinguish Common Species of Dermatophytes
This report describes the application of PCR fingerprinting for the identification of species and varieties of common dermatophytes and related fungi utilizing as a single primer the simple repetitive oligonucleotide (GACA) 4. The primer was able to amplify all the strains, producing species-specific profiles for Microsporum canis, Microsporum gypseum, Trichophyton rubrum, Trichophyton ajelloi, and Epidermophyton floccosum. Intra-specific variability was not observed for these species. Instead, three different profiles were observed in the Trichophyton mentagrophytes group.
Identification of dermatophytes by arbitrarily primed PCR
Asian Biomedicine, 2015
Background Dermatophytosis is a superficial infection caused by filamentous fungi belonging to the following three genera: Microsporum, Trichophyton and Epidermophyton. Arbitrarily primed PCR (AP-PCR) is a rapid and sensitive procedure for the diagnosis of the fungal species. Objectives To identify various dermatophyte species as rapidly and precisely as possible. Methods Fifty-two clinical dermatophyte isolates from ten species were recovered from samples obtained the Department of Medical Mycology and patients in different parts of Iran. All 52 dermatophyte isolates tested belonged to any of Trichophyton, Microsporum, or Epidermophyton genera. Four random primers, OPAA11, OPU15, OPAA17, and OPD18, were used in this study. Results The results indicated that all 10 dermatophyte species displayed distinct DNA band patterns after amplification with the random primers OPAA11 and OPU15. Nine species of dermatophytes were distinguished with the random primer OPAA17 using a different DNA ...
A comparative study on morphological versus molecular identification of dermatophyte isolates
Journal de mycologie médicale, 2015
Dermatophytes are taxonomically classified in the genera Trichophyton, Microsporum, and Epidermophyton. Pleomorphism, cultural variability, slow growth and sporulation, and the need for additional physiological tests make dermatophytes notoriously difficult to identify. The present study aimed to compare the results of morphological and molecular identification of certain groups of clinical isolates of dermatophytes with a view to evaluating the accuracy of molecular methods. For each sample, the ITS1-5.8S-ITS2 rDNA region was amplified using the primers ITS1 and ITS4. PCR products were subjected to restriction fragment length polymorphism (RFLP) analysis using the enzyme MvaI and isolate identification was performed by comparing the electrophoretic RFLP patterns with reference profiles obtained previously. Finally, paired comparative analyses of molecular and conventional methods were performed. While morphology results from routine daily reports of the laboratories indicated that ...
Journal of Clinical Microbiology, 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,
Dermatophyte identification is based on the detection of fungal elements by direct microscopy of clinical specimens combined with culture-based full identification. Phenotypic identification includes macromorphological, micromorphological, and physiological characteristics of the colonies. In the last few years, molecular approaches have been proven to be useful for identification of dermatophyte species. Objective To investigate the conventional and molecular methods used for identification of dermatophytes.
Real-time PCR approach in dermatophyte detection and Trichophyton rubrum identification
Acta Biochimica Polonica, 2015
Dermatophytes are keratinophilic molds that infect human hair, nails and skin. Diagnosis of dermatophytosis is based on morphological, serological and biochemical features. However, identification is difficult and laborious due to similarities between microorganisms. Thus, there is considerable interest to develop mycological diagnostic procedures based on molecular biology methods. In this study, fast, two-step DNA extraction method and real-time PCR was used for detection of dermatophytes DNA using pan-dermatophyte primers and identification of Trichophyton rubrum from pure cultures. The applied method allowed correct detection of all dermatophytes and correct identification of Trichophyton rubrum in less than 2 hours.
Dermatophyte and non dermatophyte fungi in Riyadh City, Saudi Arabia
Saudi Journal of Biological Sciences, 2015
Background: Dermatophytes are a scientific label for a group of three genera (Microsporum, Epidermophyton and Trichophyton) of fungus that causes skin disease in animals and humans. Conventional methods to identification of these fungi are rapid and simple but are not accurate comparing to molecular methods.