Direct detection and differentiation of causative fungi of onychomycosis by multiplex polymerase chain reaction-based assay

Li, Xiaofang; Tian, Wei; Wang, Hong; Chen, Hui; Shen, Ya; Lv, Guixia; Liu, Weida

doi:10.1684/ejd.2010.1152

Cited by 26 publications

(24 citation statements)

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“…Our results are in agreement with most previously studies [1,3,5,6,8,13,21,25]. However, in some reports, specificity was found suboptimal [6,9].…”

Section: Discussionsupporting

confidence: 93%

“…Sensitivity values reported in previous studies using different PCR methods and primers ranged between 51% and 94, 8% [1,3,5,8,20,21,31]. On the other hand, our results showed the PCR to be more sensitive as compared to mycological examination (90.5% vs 81.1%).…”

Section: Discussionmentioning

confidence: 37%

“…On the other hand, culture is time-consuming and over-growing of molds in the culture medium can prevent the development of the pathogen. Last, the sensitivity of culture is often suboptimal or low [1,5,6,12,21].…”

Section: Discussionmentioning

confidence: 99%

“…Genotypic approaches such as PCR [9,13,20,22,32,33], PCR fingertyping [13], restriction fragment length polymorphism [10,11,17,25,28,30], arbitrarily primed PCR [23], sequencing [27,28], PCR-Elisa [3], PCR-reverse line blot [4], Nested PCR [12,14], multiplex PCR [8,21,31], oligonucleotide array [9] and real-time PCR [1,5,6], are promising techniques for developing rapid and accurate diagnosis of dermatophytes by allowing to identify species and to discriminate between strains within a comparatively short period of time.…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Evaluation of Chitine synthase ( CHS1 ) polymerase chain reaction assay in diagnosis of dermatophyte onychomycosis

Dhib

Fathallah

Charfeddine

et al. 2012

Journal de Mycologie Médicale

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“…Our results are in agreement with most previously studies [1,3,5,6,8,13,21,25]. However, in some reports, specificity was found suboptimal [6,9].…”

Section: Discussionsupporting

confidence: 93%

Section: Discussionmentioning

confidence: 37%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Evaluation of Chitine synthase ( CHS1 ) polymerase chain reaction assay in diagnosis of dermatophyte onychomycosis

Dhib

Fathallah

Charfeddine

et al. 2012

Journal de Mycologie Médicale

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“…ITS sequences are increasingly used because of their high copy number, enabling amplification from a small amount of DNA, and the high degree of variation even among closely related species. [12][13][14][15][16][17] While some authors have concentrated on a pandermatophyte approach, 15,17 others attempted mainly to identify Trichophyton rubrum, as the most common infectious organism of dermatophyte infections. 13,14,16 In more recent studies, authors have tried increasingly to provide species-specific diagnostic tools, 18 especially those using real-time polymerase chain reaction (PCR).…”

mentioning

confidence: 99%

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

et al. 2015

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Evaluation of different DNA extraction methods based on steel‐bullet beating for molecular diagnosis of onychomycosis

Motamedi

Amini

Yazdanpanah

et al. 2022

Clinical Laboratory Analysis

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Background Considering increased trends toward molecular methods for detection/identification of fungi causing onychomycosis, the aim of this study is comparison three DNA extraction methods based on steel‐bullet beating to extract DNA from nail. Methods Ex ‐vivo onychomycosis model was developed using bovine hoof with Candida albicans and Aspergillus flavus. For two models, total DNA was extracted using the three different methods. In method 1, the extraction and purification were performed by steel‐bullet beating and phenol chloroform protocol, respectively. In method 2, a freezing step were applied before beating. The purification step in method 3 was carried out using a commercial kit, although DNA extraction was done similarly to method 1 in that approach. To evaluate the efficacy of each method, the extracted genomic DNA was amplified with Polymerase Chain Reaction (PCR) using Internal Transcribed Spacer (ITS) regions. Moreover, 50 nail samples were evaluated for onychomycosis using direct microscopy examination as well as PCR in order to evaluate the diagnostic efficiency of the optimal DNA extraction method. Results Regarding the desirable quality of the extracted DNA, cost effectiveness, and simplicity, method 1 could be used to extract DNA effectively. Additionally, the obtained data showed that PCR had a higher detection rate of fungal agents in the nail samples than direct microscopic examination. Conclusions This study demonstrated that the mechanical disruption of the cell wall by steel‐bullet beating is a useful and practical method to improve the quantity and quality of fungal DNA thorough the extraction process.

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Direct detection and differentiation of causative fungi of onychomycosis by multiplex polymerase chain reaction-based assay

Cited by 26 publications

References 0 publications

Evaluation of Chitine synthase ( CHS1 ) polymerase chain reaction assay in diagnosis of dermatophyte onychomycosis

Evaluation of Chitine synthase ( CHS1 ) polymerase chain reaction assay in diagnosis of dermatophyte onychomycosis

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

Evaluation of different DNA extraction methods based on steel‐bullet beating for molecular diagnosis of onychomycosis

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