Direct Identification of Dermatophyte DNA From Clinical Specimens by a Nested Polymerase Chain Reaction Assay

Yang, Cheng‐San

doi:10.1001/archderm.143.6.799

Cited by 7 publications

(7 citation statements)

References 6 publications

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“…Few studies using DNA extraction directly from clinical material without prior culture, focused mainly on identification of the infecting agent in nail specimens, have been published 21,24,32–34 . Only two recently published investigations also involved clinical specimens from patients with tinea pedis, tinea corporis, tinea manus, tinea cruris and tinea capitis 13,25 . Fortunately, the use of a commercial kit, as described in this study, provides an easy, fast and well‐standardised method for the extraction of genomic DNA.…”

Section: Discussionmentioning

confidence: 99%

“…21,24,[32][33][34] Only two recently published investigations also involved clinical specimens from patients with tinea pedis, tinea corporis, tinea manus, tinea cruris and tinea capitis. 13,25 Fortunately, the use of a commercial kit, as described in this study, provides an easy, fast and well-standardised method for the extraction of genomic DNA. The overnight Proteinase K treatment and binding of the DNA on silica gel membranes replace previously described phenol-chloroform extraction or alcohol precipitation, 32,[35][36][37] which clearly are more laborious.…”

Section: Discussionmentioning

confidence: 99%

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Direct detection of five common dermatophyte species in clinical samples using a rapid and sensitive 24-h PCR-ELISA technique open to protocol transfer

Beifuß

Bezold²,

Gottlöber³

et al. 2011

Mycoses

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Identification of dermatophytes is usually based on morphological characteristics determined by time-consuming microscopic and cultural examinations. An effective PCR-ELISA method has been developed for rapid detection of dermatophyte species directly from clinical specimens within 24 h. Isolated genomic DNA of skin scrapings and nail samples from patients with suspected dermatophyte infections is amplified with species-specific digoxigenin-labelled primers targeting the topoisomerase II gene. The subsequent ELISA procedure with biotin-labelled probes allows a sensitive and specific identification of the five common dermatophytes -Trichophyton rubrum, T. interdigitale, T. violaceum, Microsporum canis and Epidermophyton floccosum. PCR-ELISA, based on the new polyphasic species concept, was assessed using 204 microscopy-positive samples in two university mycological laboratories in Munich and Tübingen, and 316 consecutive specimens - regardless of mycological findings - in a dermatological practice laboratory in Neu-Ulm. One of the five dermatophytes was confirmed by PCR-ELISA in 163 of 204 (79.9%) of the clinical samples from the university hospitals found positive using microscopy. Culture was positive for dermatophytes in 59.8% of the same cases. A significant difference between these two methods could be demonstrated using the McNemar test (P < 0.005). Analysis of specimens from Neu-Ulm confirmed the results in a dermatological practice laboratory as 25.0% of the specimens had positive PCR results, whereas only 7.3% were positive according to culture. Direct DNA isolation from clinical specimens and the PCR-ELISA method employed in this study provide a rapid, reproducible and sensitive tool for detection and discrimination of five major dermatophytes at species level, independent of morphological and biochemical characteristics.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Direct detection of five common dermatophyte species in clinical samples using a rapid and sensitive 24-h PCR-ELISA technique open to protocol transfer

Beifuß

Bezold²,

Gottlöber³

et al. 2011

Mycoses

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“…These tests not only help in the rapid and early diagnosis of infection but also help in determining drug resistance,[ 25 ] and include:…”

Section: Diagnosis Of Dermatophytosismentioning

confidence: 99%

Management of tinea corporis, tinea cruris, and tinea pedis: A comprehensive review

Sahoo

Mahajan

2016

Indian Dermatol Online J

222

114

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The prevalence of superficial mycotic infection worldwide is 20–25% of which dermatophytes are the most common agents. Recent developments in understanding the pathophysiology of dermatophytosis have confirmed the central role of cell-mediated immunity in countering these infections. Hence, a lack of delayed hypersensitivity reaction in presence of a positive immediate hypersensitivity (IH) response to trichophytin antigen points toward the chronicity of disease. Diagnosis, though essentially clinical should be confirmed by laboratory-based investigations. Several new techniques such as polymerase chain reaction (PCR) and mass spectroscopy can help to identify the different dermatophyte strains. Management involves the use of topical antifungals in limited disease, and oral therapy is usually reserved for more extensive cases. The last few years have seen a significant rise in the incidence of chronic dermatophyte infections of skin which have proven difficult to treat. However, due to the lack of updated national or international guidelines on the management of tinea corporis, cruris, and pedis, treatment with systemic antifungals is often empirical. The present review aims to revisit this important topic and will detail the recent advances in the pathophysiology and management of tinea corporis, tinea cruris, and tinea pedia while highlighting the lack of clarity of certain management issues.

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“…Newer studies have looked at the role of PCR in the diagnosis of dermatophyte infections. This technique may be more sensitive than culture when there is a minute fungal load or a patient has been pretreated with a fungicidal drug, although high cost and poor availability discourage the broad use of PCR 31–33 . A small study by Slowinska et al.…”

Section: Diagnostic Technique and Differential Diagnosesmentioning

confidence: 99%

Tinea capitis: still an unsolved problem?

Patel

Schwartz

2011

Mycoses

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Tinea capitis is a fungal infection specifically involving the scalp and hair. It is the most common dermatophyte infection in children under 12 years of age, with a predominance in those of sub-Saharan African descent. Common signs include hair loss, scaling, erythema and impetigo-like plaques. Adults may also be affected, but to a lesser degree. The causative species are from the Microsporum and Trichophyton genera. Limited treatment options and diverse modes of transmission complicate the clinician's ability to address this disease adequately. Although dermatophytes are ubiquitous in our environment and tinea capitis is common, therapeutic options can be utilised to reduce morbidity.

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Direct Identification of Dermatophyte DNA From Clinical Specimens by a Nested Polymerase Chain Reaction Assay

Cited by 7 publications

References 6 publications

Direct detection of five common dermatophyte species in clinical samples using a rapid and sensitive 24-h PCR-ELISA technique open to protocol transfer

Direct detection of five common dermatophyte species in clinical samples using a rapid and sensitive 24-h PCR-ELISA technique open to protocol transfer

Management of tinea corporis, tinea cruris, and tinea pedis: A comprehensive review

Tinea capitis: still an unsolved problem?

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