Application of Microsatellite-Primed PCR (MSP-PCR) and PCR Melting Profile (PCR-MP) Method for Intraspecies Differentiation of Dermatophytes

Ciesielska, Anita; Kozłowska, Magdalena; Gadzalski, Marek; Worek, Mariusz; Jaworski, Adam; Stączek, Paweł

doi:10.33073/pjm-2014-038

Cited by 8 publications

(5 citation statements)

References 22 publications

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“…These techniques can also help to characterize infecting strains and monitor their occurrence and distribution [31]. Moreover, the most important investigation in the molecular epidemiology of dermatophytes is to determine whether infections are caused by the same or different strains [51]. In this aspect, disclosure of infection sources and transmission pathways in populations of humans and animals is necessary, and available techniques should allow deep genetic differentiation of strains within species, thus facilitating prompt and reliable identification of individual clones [52].…”

Section: Discussionmentioning

confidence: 99%

“…Our investigation showed a relatively high genomic diversity revealed by the MP-PCR analysis of clinical isolates of T. mentagrophytes of both human and animal origin. Although the MP-PCR method is widely described in the literature as a useful tool for the epidemiological analysis of the source of infection [31,49,51,53], it seems that it cannot be used to detect recalcitrant to treatment dermatophyte isolates. The molecular basis of terbinafine resistance is most widely described to result mostly from changes at the genome level [18,27,54,55].…”

Section: Discussionmentioning

confidence: 99%

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Population differentiation, antifungal susceptibility, and host range of Trichophyton mentagrophytes isolates causing recalcitrant infections in humans and animals

Gnat

Łagowski

Nowakiewicz

et al. 2020

Eur J Clin Microbiol Infect Dis

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The major problems in determining the causative factors of the high prevalence of dermatophytoses include the lack of a well-standardized antifungal susceptibility testing method, the low consistency of in vitro and clinical minimal inhibitory concentration values, the high genomic diversity of the population, and the unclear mechanism of pathogenicity. These factors are of particular importance when the disease is recalcitrant and relapses. Herein, we identified and characterized Trichophyton mentagrophytes isolates obtained from therapy-resistant cases in humans and animals. We used genomic diversity analysis of 17 human and 27 animal clinical isolates with the MP-PCR technique, determined their phenotypic enzymatic activity and host range, and performed antifungal susceptibility testing to currently available antifungal drugs from various chemical groups. Genomic diversity values of 35.3% and 33.3% were obtained for clinical isolates from humans and animals, respectively, yet without any relationship to the host species or antifungal drug to which resistance in therapy was revealed. The highest activity of keratinase enzymes was recorded for fox, guinea pig, and human hairs. These hosts can be considered as the main species in the host range of these isolates. A phenyl morpholine derivative, i.e. amorolfine, exhibited superior activity against strains obtained from both humans and animals with the lowest MIC50. Interestingly, high compliance of terbinafine in vitro resistance with clinical problems in the treatment with this substance was shown as well. The high resistance of dermatophytes to drugs is the main cause of the recalcitrance of the infection, whereas the other features of the fungus are less important.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Population differentiation, antifungal susceptibility, and host range of Trichophyton mentagrophytes isolates causing recalcitrant infections in humans and animals

Gnat

Łagowski

Nowakiewicz

et al. 2020

Eur J Clin Microbiol Infect Dis

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“…In the present study, we demonstrated the application of the PCR melting profile as a simple method that may be used for intraspecies differentiation of analysed strains of A. keratinophilus isolated from pellets of Corvus frugilegus . This method has been successfully used for distinguishing bacterial strains such as S. aureus or E. coli 42 and for differentiation of dermatophytes 18 , 43 . We chose the Bam HI restriction enzyme and an appropriate denaturation temperature to obtain high differentiation power with the PCR-MP method.…”

Section: Discussionmentioning

confidence: 99%

Dispersal of Aphanoascus keratinophilus by the rook Corvus frugilegus during breeding in East Poland

Kitowski

Korniłłowicz-Kowalska

Bohacz

et al. 2022

Sci Rep

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The process of dispersal of the potentially disease-causing, geophilic and keratinolytic fungal strain Aphanoascus keratinophilus (the perfect, sexual stage of Chrysosporium keratinophilum) by the rook Corvus frugilegus was studied. The source of A. keratinophilus strains was pellets of the rook, thus far not considered a carrier of this particular opportunistic pathogen. Pellets collected from breeding colonies of rooks were analysed in terms of the occurrence of keratinolytic fungi with the application of the native keratin bait method. Among the 83 rook pellets analysed, 24 (29%) were infected by keratinophilic fungi. Pure cultures of the fungi were identified to species based on traditional morphological features. Traditional mycological identification was verified by the PCR–RFLP molecular identification method as well as DNA sequencing. The obtained results showed the presence of 90 Aphanoascus keratinophilus strains, 6 Chrysosporium tropicum strains, and 3 Chrysosporium pannicola strains. The PCR melting profile (PCR-MP) method was used to identify intraspecies variations of the 90 analysed A. keratinophilus strains. The dispersal of genotypes and possible pathways of A. keratinophilus dispersal and infection via rook pellets were analysed.

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“…After onychomycosis , which, by far, is the most difficult to cure, and tinea capitis , the most prevalent superficial mycosis in the United States and Canada is tinea pedis (affecting approximately 40% of the population), followed by tinea corporis and tinea cruris 5 , 6 . Therefore, fungal skin infections, including those caused by dermatophytes, are now referred to as civilization diseases, which are a significant public health problem 7 – 9 .…”

Section: Introductionmentioning

confidence: 99%

Evaluation of the antidermatophytic activity of potassium salts of N-acylhydrazinecarbodithioates and their aminotriazole-thione derivatives

Ciesielska,

Kowalczyk,

Paneth

et al. 2024

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Nowadays, dermatophyte infections are relatively easy to cure, especially since the introduction of orally administered antifungals such as terbinafine and itraconazole. However, these drugs may cause side effects due to liver damage or their interactions with other therapeutics. Hence, the search for new effective chemotherapeutics showing antidermatophyte activity seems to be the urge of the moment. Potassium salts of N-acylhydrazinecarbodithioates are used commonly as precursors for the synthesis of biologically active compounds. Keeping that in mind, the activity of a series of five potassium N-acylhydrazinecarbodithioates (1a–e) and their aminotriazole-thione derivatives (2a–e) was evaluated against a set of pathogenic, keratinolytic fungi, such as Trichophyton ssp., Microsporum ssp. and Chrysosporium keratinophilum, but also against some Gram-positive and Gram-negative bacteria. All tested compounds were found non-toxic for L-929 and HeLa cells, with the IC30 and IC50 values assessed in the MTT assay above 128 mg/L. The compound 5-amino-3-(naphtalene-1-yl)-4,5-dihydro-1H-1,2,4-triazole-5-thione (2d) was found active against all fungal strains tested. Scanning Electron Microscopy (SEM) revealed inhibition of mycelium development of Trichophyton rubrum cultivated on nail fragments and treated with 2d 24 h after infection with fungal spores. Transmission Electron Microscopy (TEM) observation of mycelium treated with 2d showed ultrastructural changes in the morphology of germinated spores. Finally, the RNA-seq analysis indicated that a broad spectrum of genes responded to stress induced by the 2d compound. In conclusion, the results confirm the potential of N-acylhydrazinecarbodithioate derivatives for future use as promising leads for new antidermatophyte agents development.

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Application of Microsatellite-Primed PCR (MSP-PCR) and PCR Melting Profile (PCR-MP) Method for Intraspecies Differentiation of Dermatophytes

Cited by 8 publications

References 22 publications

Population differentiation, antifungal susceptibility, and host range of Trichophyton mentagrophytes isolates causing recalcitrant infections in humans and animals

Population differentiation, antifungal susceptibility, and host range of Trichophyton mentagrophytes isolates causing recalcitrant infections in humans and animals

Dispersal of Aphanoascus keratinophilus by the rook Corvus frugilegus during breeding in East Poland

Evaluation of the antidermatophytic activity of potassium salts of N-acylhydrazinecarbodithioates and their aminotriazole-thione derivatives

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