Development and validation of a real-time multiplex PCR assay for the detection of dermatophytes and Fusarium spp.

Koo, Seok Hwee; Teoh, Yee Leng; Koh, Wei Liang; Ochi, Harumi; Tan, Sher Kye; Sim, Diana Miao Fang; Jiang, Boran; Tan, Ai Ling; Tan, Thean Yen; Lim, Su Ping Regina

doi:10.1099/jmm.0.001082

Cited by 14 publications

(7 citation statements)

References 25 publications

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“…This variant of conventional PCR increases the efficiency of detection and reduces the risk of false-negative results. Highlighted the efficiency, sensitivity, and specificity of multiplex PCR for a faster diagnosis of onychomycosis caused by dermatophytes and by Fusarium spp [59]. Similarly, recommended the use of multiplex PCR for the diagnosis of dermatophytic onychomycosis in cases with a negative culture or when the culture is contaminated with fast-growing fungi, a fact that renders the identification of the causal agent problematic [60].…”

Section: Culture Microscopy Histology and Molecular Assays Based Onmentioning

confidence: 99%

Epidemiology and Diagnostic Perspectives of Dermatophytoses

Petrucelli

Abreu

Cantelli

et al. 2020

JoF

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Dermatophytoses affect about 25% of the world population, and the filamentous fungus Trichophyton rubrum is the main causative agent of this group of diseases. Dermatomycoses are caused by pathogenic fungi that generally trigger superficial infections and that feed on keratinized substrates such as skin, hair, and nails. However, there are an increasing number of reports describing dermatophytes that invade deep layers such as the dermis and hypodermis and that can cause deep infections in diabetic and immunocompromised patients, as well as in individuals with immunodeficiency. Despite the high incidence and importance of dermatophytes in clinical mycology, the diagnosis of this type of infection is not always accurate. The conventional methods most commonly used for mycological diagnosis are based on the identification of microbiological and biochemical features. However, in view of the limitations of these conventional methods, molecular diagnostic techniques are increasingly being used because of their higher sensitivity, specificity and rapidity and have become more accessible. The most widely used molecular techniques are conventional PCR, quantitative PCR, multiplex PCR, nested, PCR, PCR-RFLP, and PCR-ELISA. Another promising technique for the identification of microorganisms is the analysis of protein profiles by MALDI-TOF MS. Molecular techniques are promising but it is necessary to improve the quality and availability of the information in genomic and proteomic databases in order to streamline the use of bioinformatics in the identification of dermatophytes of clinical interest.

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Section: Culture Microscopy Histology and Molecular Assays Based Onmentioning

confidence: 99%

Epidemiology and Diagnostic Perspectives of Dermatophytoses

Petrucelli

Abreu

Cantelli

et al. 2020

JoF

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“…The literature reports numerous PCR studies conducted on patients with suspected onychomycosis 6‐18 . Most studies use home‐made PCR techniques which are time‐consuming and difficult to perform without a dedicated laboratory: they are useful in epidemiological studies of onychomycosis, but not in clinical practice.…”

Section: Discussionmentioning

confidence: 99%

Real‐life applicability of the Euroarray dermatomycosis kit in the diagnosis of onychomycosis

Trave

Cozzani

Canepa

et al. 2022

Mycoses

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Background Traditionally, KOH microscopy and fungal culture are the two preferred tests as gold standard for diagnosis of onychomycosis. Recently, other diagnostic methods have been developed to improve the microbiological diagnosis. The EUROArray dermatomycosis kit is a PCR‐based microarray test system for the detection and direct identification of species that are most frequently involved in skin and nail infections. Objectives Our primary aim was to evaluate the real‐life applicability of the EUROArray dermatomycosis kit in the diagnosis of onychomycoses. In addition, we compared the aetiology of onychomycoses found in our patients with those described in the literature. Patients/Methods We prospectively studied consecutive 100 patients with suspected onychomycoses. Samples of suspect toenails were taken as part of routine medical management. Nail specimens were evaluated by means of three diagnostic methods: KOH preparation, culture and EUROArray dermatomycosis kit. Results Onychomycosis was diagnosed in 47/100 patients who proved positive on at least one reference diagnostic test and in 49/100 patients who proved positive on PCR. The combination of microscopy and PCR had better sensitivity than microscopy (p = .0397), fungal culture (p = .0061) and PCR alone (p = .0117). Moulds were more frequently positive in culture than in PCR (p = .033). Dermatophytes proved positive more frequent than moulds and yeasts in both culture and PCR; in particular, Trichophyton interdigitale was the most frequent pathogen. Conclusions In conclusion, introducing EUROArray dermatomycosis kit into the diagnostic algorithm of onychomycosis increases the sensitivity of direct microscopy and yields more rapid results than culture.

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“…Due to the similarity in fungal beta diversity between a subclass of OM patients with healthy controls, ITS2 sequencing, followed by taxonomic annotation, might currently not be suited for the diagnosis of OM. Thus, diagnosis of OM still seems to be a domain of conventional microscopy and culture, which may be partially replaced by sequencing-based multiplex assays 6 , 32 . However, ITS sequencing, in addition to conventional/molecular OM diagnosis, may provide significant clues regarding the underlying mechanics of infection and disease progression, i.e., whether or not additional host- and/or environmental factors need to be considered for treatment and prevention.…”

Section: Discussionmentioning

confidence: 99%

Biodiversity of mycobial communities in health and onychomycosis

Olbrich

Ernst

Beltsiou

et al. 2022

Sci Rep

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Onychomycosis (OM) is a common fungal nail infection. Based on the rich mycobial diversity in healthy toenails, we speculated that this is lost in OM due to the predominance of a single pathogen. We used next generation sequencing to obtain insights into the biodiversity of fungal communities in both healthy individuals and OM patients. By sequencing, a total of 338 operational-taxonomic units were found in OM patients and healthy controls. Interestingly, a classifier distinguished three distinct subsets: healthy controls and two groups within OM patients with either a low or high abundance of Trichophyton. Diversity per sample was decreased in controls compared to cases with low Trichophyton abundance (LTA), while cases with a high Trichophyton abundance (HTA) showed a lower diversity. Variation of mycobial communities between the samples showed shifts in the community structure between cases and controls—mainly driven by HTA cases. Indeed, LTA cases had a fungal β-diversity undistinguishable from that of healthy controls. Collectively, our data provides an in-depth characterization of fungal diversity in health and OM. Our findings also suggest that onychomycosis develops either through pathogen-driven mechanisms, i.e., in HTA cases, or through host and/or environmental factors, i.e., in cases with a low Trichophyton abundance.

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Development and validation of a real-time multiplex PCR assay for the detection of dermatophytes and Fusarium spp.

Cited by 14 publications

References 25 publications

Epidemiology and Diagnostic Perspectives of Dermatophytoses

Epidemiology and Diagnostic Perspectives of Dermatophytoses

Real‐life applicability of the Euroarray dermatomycosis kit in the diagnosis of onychomycosis

Biodiversity of mycobial communities in health and onychomycosis

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