Diagnosis of common dermatophyte infections by a novel multiplex real-time polymerase chain reaction detection/identification scheme

Arabatzis, Michael; Coppenraet, L.E.S. Bruijnesteijn van; Kuijper, Ed J.; Hoog, G.S. de; Lavrijsen, A.P.M.; Templeton, Kate; Raaij-Helmer, Elizabeth M.H. van der; Velegraki, Aristea; Gräser, Yvonne; Summerbell, Richard C.

doi:10.1111/j.1365-2133.2007.08100.x

Cited by 116 publications

(131 citation statements)

References 27 publications

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“…Secondly, we further augmented the performances of this type of test by processing a large quantity of material collected by skin scrapings from each animal. Finally, results obtained with microscopic examination were confirmed by real-time PCR, which is known to possess a sensitivity and a specificity comparable or even superior to that of culture [12]. The PCR employed has been proven to detect different species of Trichophyton [12], so that the exact fungal identification was based on the morphological features of fungal elements visualized at the microscopic level.…”

Section: Discussionmentioning

confidence: 88%

“…A minor groove binder (MGB) Taqman probe labeled with a VIC reporter dye at the 5'-end (Life Technologies, Monza, Italy) was used. This set of primers and probe is known to detect all pathogenic species of Trichophyton, including T. verrucosum [12]. Amplification reactions contained 10 µL DNA extract, 25 µL of 2 X Taqman Universal Master Mix (Life Technologies, Monza, Italy), 0.4 µmol L -1 of each primer and 0.1 µmol L -1 of the probe, with nuclease-free water up to a final volume of 50 µL.…”

Section: Laboratory Proceduresmentioning

confidence: 99%

“…Moreover, most T. verrucosum isolates require thiamine, or thiamine and inositol, though some autotrophic variants that do not require an exogenous source of vitamins, have been described [3]. Recently, a number of molecular methods have also been proposed as alternative tools to detect dermatophytes, including T. verrucosum directly from clinical samples [12].…”

Section: Introductionmentioning

confidence: 99%

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Trichophyton verrucosum infection in livestock in the Chitral district of Pakistan

Hameed

Riaz

Nawaz

et al. 2017

J Infect Dev Ctries

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Introduction: Trichophyton verrucosum belongs to the dermatophyte fungi, closely related organisms that cause skin infections in animals and humans. T. verrucosum infection has been reported in livestock and people in different countries from all continents. Human cases have been reported in different areas of Pakistan, but there is little information about the animal source of the fungus. Methodology: Dermatological specimens collected in the Chitral district of Pakistan for a study on mange in livestock were retrospectively analyzed for the presence of T. verrucosum. In total, 5,873 animals (1,087 cows, 2,033 goats, and 2,753 sheep) were screened for evidence of dermatological lesions during two surveys performed in the summer and winter seasons. Skin scrapings collected from animals with lesions were analyzed by direct microscopic examination after digestion in sodium hydroxide and a real-time polymerase chain reaction (PCR) targeting pathogenic Trichophyton species. Results: At microscopy, samples from 18 cows (1.6%), 3 sheep (0.1%), and 4 goats (0.2%) were positive for fungal elements consistent with T. verrucosum. PCR confirmed the microscopy results. The prevalence was lower than that reported in other countries in intensive breeding farms. Results agree with the literature regarding factors affecting T. verrucosum diffusion, i.e., infection was more prevalent in cattle, especially in younger animals during the winter season. Conclusions: This study reports, for the first time, the presence of T. verrucosum in livestock in Pakistan. A better knowledge of the animal role in the spread of this fungus may allow the adoption of more efficient control measures and prophylaxis.

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Section: Discussionmentioning

confidence: 88%

Section: Laboratory Proceduresmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Trichophyton verrucosum infection in livestock in the Chitral district of Pakistan

Hameed

Riaz

Nawaz

et al. 2017

J Infect Dev Ctries

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“…Currently, Fungus responsible for the infection can be identified with great precision by modern technologies like-PCR and Nucleic acid based tests. 5,6 Present hospital based observational study was carried out to find out the prevalence of dermatophytosis, its clinical presentation and species of fungi responsible for the disease in Central Nepal…”

Section: Introductionmentioning

confidence: 99%

“Epizoonosis of Dermatophytosis”: A Clinico - Mycological Study of Dermatophytic Infections in Central Nepal

Mathur

Kedia

Ghimire

2012

Kathmandu Univ. Med. J.

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“…However, outside factors such as temperature variation, medium and chemotherapy, can greatly influence the phenotypic characteristic and consequently can make the identification more difficult (Weitzman and Summerbell, 1995;Faggi et al, 2001;Kanbe, 2008;De Baere et al, 2010). Recently, molecular marker approaches, such as nested-polymerase chain reaction (PCR) (Verrier et al, 2013), random amplified polymorphic DNA (RAPD)-PCR (Kim et al, 2001;Baeza et al, 2006;Leibner-Ciszak et al, 2010;Dobrowolska et al, 2011;Spesso et al, 2013), inter-simple sequence repeat (ISSR)-PCR (Cano et al, 2005;Khosrav et al, 2012), PCR-restriction fragment length polymorphism (RFLP) (Yang et al, 2008;RezaeiMatehkolaei et al, 2012;Samuel et al, 2013), real time PCR (Bergmans et al, 2010) and multiplex PCR assay (Arabatzis et al, 2007;Kim et al, 2011) and others have been adapted for detection of dermatophytes from clinical specimens.…”

Section: Introductionmentioning

confidence: 99%

Genetic relationships and isozyme profile of dermatophytes and Candida strains from Egypt and Libya

Abdel-Fatah¹,

Moharram²,

Moubasher³

et al. 2013

Afr. J. Biotechnol.

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Three molecular techniques random amplified polymorphic DNA (RAPD), inter-simple sequence repeat (ISSR) and restriction fragment length polymorphism (RFLP) were employed for identification and to study the genetic relationship among six species of dermatophytes and three species of yeasts isolated from Egyptian and Libyan patients with skin mycosis. Each species was represented by two isolates, one from Egyptian patients and the second from Libyan. RAPD in which four random 10-mer primers and two ISSR primers were used to amplify the DNA fragments of target fungi and RFLP in which two universal primers (ITS1 and ITS4) were used to amplify the internal transcribed spacer (ITS) regions of the ribosomal (rRNA) gene in fungal isolates followed by digestion with HinfI and HaeIII endonucleases was carried out. Three molecular marker techniques showed considerable potential for identifying and discriminating dermatophytes and Candida species and the achieved results confirmed identification based on conventional morphological methods. Results of RAPD and ISSR markers revealed 78.7% genetic similarity (GS) between Microsporum canis and other tested fungi reflecting a relatively longer genetic distance from other isolates of dermatophytes and yeasts. Candida krusei and Candida tropicalis were closely related showing 93.3% GS. C. albicans showed 90.9% similarity with other species of Candida. Epidermophyton floccosum was easily separated from all Trichophyton species showing 87.3% similarity. Unique bands were displayed by certain fungi and can be taken as a positive marker for isolate identification and discrimination. RFLP technique revealed differences in the number (1 to 5) and size (8 to 378 base pairs) of DNA fragments depending on the fungal isolate and restriction enzyme used. Within each fungal species, different isolates of dermatophytes and Candida from Egypt and Libya showed close relationship. Seven isozyme systems namely esterase, peroxidase, malate dehydrogenase, acid phosphatase, glutamate-oxalo-acetate transaminase, Urease and protease were studied to detect the gene expression and genetic variability among the different isolates of dermatophytes and Candida.

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Diagnosis of common dermatophyte infections by a novel multiplex real-time polymerase chain reaction detection/identification scheme

Abstract: This highly sensitive assay also proved to have high positive and negative predictive values (95.7% and 100%), facilitating the accurate, rapid diagnosis conducive to targeted rather than empirical therapy for dermatophytoses.

Cited by 116 publications

References 27 publications

Trichophyton verrucosum infection in livestock in the Chitral district of Pakistan

Trichophyton verrucosum infection in livestock in the Chitral district of Pakistan

“Epizoonosis of Dermatophytosis”: A Clinico - Mycological Study of Dermatophytic Infections in Central Nepal

Genetic relationships and isozyme profile of dermatophytes and Candida strains from Egypt and Libya

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