Species identification and strain differentiation of dermatophyte fungi using polymerase chain reaction amplification and restriction enzyme analysis

Shin, Jang-Hyun; Sung, Jung-Hyun; Park, Sang-Jin; Kim, Jeong Aee; Lee, Joo Heung; Lee, Dong‐Youn; Lee, Eil-Soo; Yang, Jun‐Mo

doi:10.1067/mjd.2003.491

Cited by 54 publications

(39 citation statements)

References 12 publications

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“…Introduction of a PCR-based methodology would increase specificity, simplicity, and speed and potentially even reduce cost. For studies on species identification and typing, PCR (7,11), PCR fingerprinting (4,12), random amplification of polymorphic DNA (14,17), PCR and restriction fragment length polymorphism analysis (12,24), and arbitrarily primed PCR (16,17) have all been applied. The main targets have been the following genes or DNA fragments: the ribosomal DNA region, DNA topoisomerase II genes, and the chitin synthase gene (11,13).…”

mentioning

confidence: 99%

Five-Hour Diagnosis of Dermatophyte Nail Infections with Specific Detection ofTrichophyton rubrum

2007

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A rapid two-step DNA extraction method and a multiplex PCR for the detection of dermatophytes in general and Trichophyton rubrum specifically were developed and evaluated with DNA extracted from pure cultures and from clinically diseased nails. DNA from the following dermatophytes was used: Epidermophyton floccosum, Microsporum audouinii, Microsporum canis, Microsporum gypseum, Microsporum nanum, Trichophyton mentagrophytes, Trichophyton rubrum, Trichophyton schoenleinii, Trichophyton soudanense, Trichophyton terrestre, Trichophyton tonsurans, Trichophyton verrucosum, and Trichophyton violaceum. Human DNA and DNA from the following nondermatophyte fungi were included as controls: Alternaria, Aspergillus niger, Candida albicans, Candida glabrata, Candida krusei, Malassezia furfur, Saccharomyces cerevisiae, and Scopulariopsis brevicaulis. A total of 118 nail samples received for routine microscopy and culture for dermatophytes were subsequently tested by the two PCRs separately and in a multiplex format. Using DNA extracted from pure cultures and the pan-dermatophyte PCR, the T. rubrum-specific PCR sequentially and in a multiplex format correctly detected all dermatophytes and additionally correctly identified T. rubrum. Comparison of the traditional diagnostic evaluation (microscopy and culture) of nail samples with PCR on DNA directly extracted from the nails showed excellent agreement between PCR and microscopy, but the number of samples with dermatophyte species identification was increased considerably from 22.9% to 41.5%, mainly due to the identification of T. rubrum by PCR in microscopy-positive but culture-negative samples. In conclusion, this 5-hour diagnostic test was shown to increase not only the speed but also the sensitivity of investigation for nail dermatophytosis.

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mentioning

confidence: 99%

Five-Hour Diagnosis of Dermatophyte Nail Infections with Specific Detection ofTrichophyton rubrum

2007

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“…Es gilt,eine genetisch durchführbare Artbestimmung zu entwickeln,deren Ergebnisse zumindest dort mit denjenigen konventioneller Methoden kompatibel sind, wo diese zu zuverlässigen und auch plausiblen Artdefinitionen in Übereinstim-mung mit den klassischen taxonomischen Richtlinien geführt haben. Zurzeit werden solche neuen Einteilungen entwickelt, wobei die Bestimmung der mit verschiedenen Primern erhaltenen zufallsamplifizierten polymorphen DNA-Abschnitte und das PCR-Fingerprinting genutzt werden.Analysiert werden vor allem die "internal transcribed spacer"-Regionen der rDNA, die Sequenzen bestimmter Gene und des "nontranscribed spacer" der rRNA [12,13,14,15,16,17,18,23] Bis eine genetische Differenzierung sämtlicher Arten und Varianten von Dermatophyten sicher und routinemäßig durchführbar ist,wird es vermutlich noch etwas dauern.Zuvor könnte es in der dermatomykologischen Diagnostik möglich werden, mittels PCR Dermatophyten-DNA rasch in Untersuchungsmaterialien nachzuweisen. Damit ließe sich immerhin eine Dermatophytose als solche schnell erkennen und gegen andere Pilzinfektionen abgrenzen.…”

Section: Genetische Untersuchungenunclassified

Bew�hrte und neue Verfahren zur Differenzierung von Dermatophyten

Brasch

2004

Der Hautarzt

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Although dermatophytoses can often be diagnosed clinically, the identification of the dermatophyte species causing the disease gives additional relevant information. Species identification is necessary to survey the current epidemiologic situation, and it is also needed in each individual case to trace back the path of infection, thereby to avoid reinfection, and to optimize the therapy. Classical methods to differentiate dermatophytes are based on the assessment of their macroscopic and microscopic morphology and on the observation of physiologic properties. For scientific taxonomic purposes mating experiments are indispensable. In recent years, DNA analysis has been refined to make a genetic-based differentiation of dermatophyte species possible, although such methods are not available for routine practice. No elaborate technical equipment is needed to differentiate dermatophytes in the office. The investigator's mycological expertise, however, is of crucial importance. Mycological courses and dermato-mycological quality control are appropriate means to ensure that dermatologists retain their competence in treating patients with dermatophytoses.

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“…El examen por microscopia de piel y uñas suele ser suficiente para el diagnóstico de una infección fúngica 22 pero no nos da identificación de género o especie de ahí que no se pueda diferenciar de manera segura entre los dermatofitos y otros hongos 5,6 . La posterior identificación de especies se realiza mediante un cultivo e identificación morfológica de las colonias de hongos 24 . El cultivo, sin embargo, es negativo en un 40% de los casos positivos por microscopia y además consume demasiado tiempo debido al lento crecimiento y esporulación y necesidad de más exámenes fisiológi-cos. El tiempo necesario para una identificación de especies puede variar desde 10-15 días hasta 3-4 semanas 28 .…”

Section: Introductionunclassified

Diagnóstico de infecciones por dermatofitos en uñas con detección rápida específica de Trichophyton rubrum

Riaño¹,

Arocas

Avilés

et al. 2011

Rev. Int. Cienc. Podol.

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Species identification and strain differentiation of dermatophyte fungi using polymerase chain reaction amplification and restriction enzyme analysis

Cited by 54 publications

References 12 publications

Five-Hour Diagnosis of Dermatophyte Nail Infections with Specific Detection ofTrichophyton rubrum

Five-Hour Diagnosis of Dermatophyte Nail Infections with Specific Detection ofTrichophyton rubrum

Bew�hrte und neue Verfahren zur Differenzierung von Dermatophyten

Diagnóstico de infecciones por dermatofitos en uñas con detección rápida específica de Trichophyton rubrum

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