Species identification of dermatophytes in paraffin-embedded biopsies with a new polymerase chain reaction assay targeting the internal transcribed spacer 2 region and comparison with histopathological features

Eckert, John; Ertas, Beyhan; Falk, Thomas; Metze, Dieter; Böer‐Auer, Almut

doi:10.1111/bjd.14281

Cited by 18 publications

(16 citation statements)

References 27 publications

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“…However, it is impossible to determine the fungal genus or species based on the morphology of hyphae and spores in the tissue. Some studies have shown that fungal structures detected by histology may possibly also be identified and differentiated by NATs on formalin-fixed and paraffin-embedded nail and skin tissue (e. g., pathogenic dermatophyte species) [4].…”

Section: Confirmation and Pathogen Identification (Species) In Case Omentioning

confidence: 99%

Molecular diagnosis of skin infections using paraffin‐embedded tissue – review and interdisciplinary consensus

Sunderkötter

Becker

Kutzner

et al. 2018

J Deutsche Derma Gesell

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Summary Nucleic acid amplification techniques (NATs), such as PCR, are highly sensitive and specific methods that have become valuable supplements to culture and serology in the diagnosis of infectious disorders. However, especially when using formalin‐fixed and paraffin‐embedded tissue, these techniques are associated with both false‐negative and false‐positive results, a pitfall that is frequently misjudged. Representatives of the German Society of Hygiene and Microbiology (DGHM) and the German Society of Dermatology (DDG) therefore set out to develop a consensus – in the form of a review article – on the appropriate indications for NATs using paraffin‐embedded tissue, its contraindications, and the key points to be considered in the pre‐ and post‐analytical phase. Given that fresh, naive tissue is preferably to be used in the workup of a suspected infection, PCR analysis on paraffin sections represents an exception. The latter may be considered if an infection is suspected at a later point in time and fresh tissue has not been preserved or can no longer be obtained. Potential indications include confirmation of histologically suspected infections with Leishmania spp., Bartonella spp., Rickettsia spp., or in case of ecthyma contagiosum. Infections with, for example, mycobacteria or RNA viruses, on the other hand, are not considered useful indications for NATs using paraffin sections. In order to avoid misinterpretation of test results, it is essential that laboratory reports on NATs using paraffin‐embedded tissue contain information on the indication/diagnostic circumstances, the required and chosen pre‐analytical steps, the limitations of the method, and on diagnostic alternatives.

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Section: Confirmation and Pathogen Identification (Species) In Case Omentioning

confidence: 99%

Molecular diagnosis of skin infections using paraffin‐embedded tissue – review and interdisciplinary consensus

Sunderkötter

Becker

Kutzner

et al. 2018

J Deutsche Derma Gesell

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“…Eckert et al used a new PCR assay targeting the ITS 2 region for identification of dermatophytes in paraffin‐embedded biopsies. They compared the results with histopathological investigation of the skin tissue.…”

Section: Discussionmentioning

confidence: 99%

Deep facial mycosis due toTrichophyton verrucosum—molecular genetic identification of the dermatophyte in paraffin‐embedded tissue—case report and review of the literature

Wollina

Hansel

Uhrlaß³

et al. 2017

Mycoses

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Deep trichophytosis is relatively uncommon. The infection of the bearded area is also known as sycosis barbae or tinea barbae and can be caused by various fungal species, most often zoophilic fungi. We report on an 80-year-old male patient with severe sycosis barbae who had no animal contact and was treated with systemic antibiosis without improvement. Microbial and mycological investigations using swabs from oozing lesions revealed Staphylococcus haemolyticus and Candida parapsilosis. Histology demonstrated fungal elements in hair follicles. Paraffin-embedded material was subjected to further mycological analysis. For molecular diagnostics DNA was prepared from paraffin sections for real-time polymerase chain reaction (RT-PCR). For sequencing, DNA was isolated from paraffin-embedded skin tissue and the ITS region of the rDNA was selected. Sequencing of the ITS2 region of rRNA revealed a 100% accordance with Trichophyton (T.) verrucosum. Treatment with oral terbinafine achieved a complete remission. Sycosis barbae is an important differential diagnosis for infections of the bearded area. Nucleic acid amplification techniques (NAAT) are more and more used for direct examination of dermatophytes in clinical samples, eg T. verrucosum. NAAT are also used as culture confirmation tests for identification of rare dermatophytes like T. verrucosum. Today, singleplex and multiplex quantitative real-time PCR (qRT-PCR) assays for the detection of the most common dermatophytes including T. verrucosum in clinical specimens are available. Recently, an ITS2 PCR assay has been successfully used for direct detection of T. verrucosum in paraffin-embedded formalin-fixed skin tissue. The PCR is fast and highly specific. The sensitivity of direct molecular detection of the dermatophytes both in native clinical material, and in paraffin-embedded skin tissue can been increased.

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“…However, we could not cultivate or isolate any species. We therefore attempted molecular identification of formalin‐fixed, paraffin‐embedded (FFPE) biopsy specimens . Fungal DNA was extracted from five thin‐sectioned FFPE tissue samples, each section approximately 10 μm, using a QIAamp DNA FFPE Tissue Kit (catalog no.…”

Section: Case Reportmentioning

confidence: 99%

Revival of favus in Japan caused by Trichophyton schoenleinii

Iwasa

Ogawa

Azukizawa

et al. 2019

The Journal of Dermatology

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Favus is a type of dermatophytosis known to produce yellow scutula around hair follicles. Most cases of this disease worldwide are infections of Trichophyton schoenleinii. Favus has rarely been reported in Japan throughout the last four decades, and T. schoenleinii has not been clinically isolated in any case during the period. Here, we report a case of favus of vellus hair observed in a 63‐year‐old Japanese woman. Fungal culture showed negative; however, we detected fungal elements in the crust and hair bulbs by Grocott staining. Pathogenic fungi were identified as T. schoenleinii by polymerase chain reaction‐based DNA sequencing, targeting the internal transcribed spacer regions of the rRNA gene using the formalin‐fixed, paraffin‐embedded tissue sample. She was successfully treated with p.o. administration of terbinafine and topical application of luliconazole cream.

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Species identification of dermatophytes in paraffin-embedded biopsies with a new polymerase chain reaction assay targeting the internal transcribed spacer 2 region and comparison with histopathological features

Abstract: Molecular species identification of dermatophytes via ITS2 PCR can easily be implemented in a routine dermatopathology setting. It is fast and highly specific and improves the sensitivity of histopathological diagnosis of dermatophytosis.

Cited by 18 publications

References 27 publications

Molecular diagnosis of skin infections using paraffin‐embedded tissue – review and interdisciplinary consensus

Molecular diagnosis of skin infections using paraffin‐embedded tissue – review and interdisciplinary consensus

Deep facial mycosis due toTrichophyton verrucosum—molecular genetic identification of the dermatophyte in paraffin‐embedded tissue—case report and review of the literature

Revival of favus in Japan caused by Trichophyton schoenleinii

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