Development and Evaluation of a Novel Real-Time PCR for Pan-Dermatophyte Detection in Nail Specimens

Gong, Jie; Ran, Menglong; Wang, Xiaowen; Wan, Zhe; Li, Ruoyu

doi:10.1007/s11046-015-9915-0

Cited by 13 publications

(10 citation statements)

References 21 publications

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“…The positive and negative controls in all experiment runs ruled out the possibility of experimental error. The sensitivity obtained for the real‐time PCR in our study (87.5%) was lower than observed in previous studies, in which the sensitivity ranged from 90.4% to 100% …”

Section: Discussioncontrasting

confidence: 91%

Clinical evaluation of β‐tubulin real‐time PCR for rapid diagnosis of dermatophytosis, a comparison with mycological methods

Motamedi

Mirhendi

Zomorodian

et al. 2017

Mycoses

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Following our previous report on evaluation of the beta tubulin real-time PCR for detection of dermatophytosis, this study aimed to compare the real-time PCR assay with conventional methods for the clinical assessment of its diagnostic performance. Samples from a total of 853 patients with suspected dermatophyte lesions were subjected to direct examination (all samples), culture (499 samples) and real-time PCR (all samples). Fungal DNA was extracted directly from clinical samples using a conical steel bullet, followed by purification with a commercial kit and subjected to the Taq-Man probe-based real-time PCR. The study showed that among the 499 specimens for which all three methods were used, 156 (31.2%), 128 (25.6%) and 205 (41.0%) were found to be positive by direct microscopy, culture and real-time PCR respectively. Real-time PCR significantly increased the detection rate of dermatophytes compared with microscopy (288 vs 229) with 87% concordance between the two methods. The sensitivity, specificity, positive predictive value, and negative predictive value of the real-time PCR was 87.5%, 85%, 66.5% and 95.2% respectively. Although real-time PCR performed better on skin than on nail samples, it should not yet fully replace conventional diagnosis.

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Section: Discussioncontrasting

confidence: 91%

Clinical evaluation of β‐tubulin real‐time PCR for rapid diagnosis of dermatophytosis, a comparison with mycological methods

Motamedi

Mirhendi

Zomorodian

et al. 2017

Mycoses

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“…Within this context, studies suggest that real-time PCR can be applied successfully for the diagnosis of dermatophytosis with much higher precision compared to classical diagnostic methods [48]. Furthermore, evaluating dermatophytes in nail samples, [66] showed that real-time PCR detected the presence of dermatophytes in samples that tested negative by conventional PCR, suggesting a higher diagnostic sensitivity of the former.…”

Section: Culture Microscopy Histology and Molecular Assays Based Onmentioning

confidence: 99%

“…In an attempt to increase the diagnostic response, real-time PCR can be performed as described by [67], who indicates multiplex RT-PCR as the fastest and most efficient method for the identification of dermatophyte species in clinical samples, or using pan-dermatophyte primers as described by [66] for the diagnosis of dermatophytes in nail samples. Commercial kits designed to facilitate the identification of dermatophytes by real-time PCR have also been developed.…”

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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“…So, in the absence of a real gold standard, it is more reasonable to use some statistical methods such as LCA, which gives a possibility to calculate not only sensitivities but also specificities. In two studies, PCR was compared to direct microscopic examination and culture with LCA, and both its sensitivity and specificity were found to be over 80% …”

Section: Discussionmentioning

confidence: 99%

“…However, there is no real gold standard to calculate their sensitivities and specificities. So, latent class analysis (LCA) has been used to determine the value of PCR in the diagnosis of onychomycosis in comparison to direct microscopic examination and culture …”

Section: Introductionmentioning

confidence: 99%

Diagnostic values of KOH examination, histological examination, and culture for onychomycosis: a latent class analysis

et al. 2018

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Background In the absence of a real gold standard, comparative studies are still done on diagnostic methods for onychomycosis. There are only a few attempts using latent class analysis to determine the value of polymerase chain reaction in comparison to conventional methods. We aimed to determine the value of histological examination in such a way for the diagnosis of onychomycosis.Methods Potassium hydroxide mount (KOH), culture and histological examination with periodic acid-Schiff (PAS), and Gomori's methenamine silver (GMS) stains were done in 106 patients having clinically suspected toenail onychomycosis.Results KOH was positive in 74% of the patients; culture in 14%; PAS in 30%; and GMS in 66%. According to the results of the latent class analysis, culture and PAS were highly specific but poorly sensitive; KOH, highly sensitive but poorly specific; and GMS, both highly sensitive and specific.Conclusions Based on these results, we have proposed KOH as a screening test and GMS as a confirmatory test for the diagnosis of onychomycosis in our own practice.However, since positivity rates of different diagnostic methods vary widely in different centers, it is more suitable that every center should determine their own diagnostic strategy by evaluating their own results with latent class analysis.

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Development and Evaluation of a Novel Real-Time PCR for Pan-Dermatophyte Detection in Nail Specimens

Cited by 13 publications

References 21 publications

Clinical evaluation of β‐tubulin real‐time PCR for rapid diagnosis of dermatophytosis, a comparison with mycological methods

Clinical evaluation of β‐tubulin real‐time PCR for rapid diagnosis of dermatophytosis, a comparison with mycological methods

Epidemiology and Diagnostic Perspectives of Dermatophytoses

Diagnostic values of KOH examination, histological examination, and culture for onychomycosis: a latent class analysis

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