Real-time PCR approach in dermatophyte detection and Trichophyton rubrum identification

Abstract

Dermatophytes are keratinophilic molds that infect human hair, nails and skin. Diagnosis of dermatophytosis is based on morphological, serological and biochemical features. However, identification is difficult and laborious due to similarities between microorganisms. Thus, there is considerable interest to develop mycological diagnostic procedures based on molecular biology methods. In this study, fast, two-step DNA extraction method and real-time PCR was used for detection of dermatophytes DNA using pan-dermatophyte primers and identification of Trichophyton rubrum from pure cultures. The applied method allowed correct detection of all dermatophytes and correct identification of Trichophyton rubrum in less than 2 hours.

PCR and real‐time PCR approaches to the identification of Arthroderma otae species Microsporum canis and Microsporum audouinii/Microsporum ferrugineum

“…The species were identified based on their macro‐ and micromorphology. In order to verify whether the isolates were dermatophytes, pan‐dermatphyte real‐time PCR was performed as described previously . Additionally, 13 non‐dermatopyhte isolates and human DNA samples from the collection of the Department of Molecular Biotechnology and Microbiology at the Gdańsk University of Technology (Poland) were included as controls.…”

“…In previous studies, we have described diagnostic PCR tests that enabled the identification of infections caused by Trichophyton ( T . ), as well as those due to M. audouinii and M. canis . Additionally, we provided a PCR test for the identification of M. canis .…”

“…In previous studies, we have described diagnostic PCR tests that enabled the identification of infections caused by Trichophyton (T.), as well as those due to M. audouinii and M. canis. 6,7 Additionally, we provided a PCR test for the identification of M. canis. 8 In this paper, we present alternative traditional and real-time PCR methods for the identification of both anthropophilic and zoophilic members of the A. otae complex using DNA extracted from pure cultures by a rapid procedure described previously.…”

Performance of the Real Fungus-ID kit based on multiplex RT-PCR assay for the rapid detection and identification of Trichophyton spp. and Microsporum spp. in clinical specimens with suspected dermatophyte infection

“…; Kobylak et al . ). Among these, PCR‐based analysis has become the most reliable method for the identification of pathogenic fungi from clinical specimens (Kondori et al .…”

“…However, culture methods are timeconsuming (2-3 weeks) due to slow growth and the need for additional biochemical testing (Sato et al 2012). In order to overcome the limitations of conventional methods, many molecular diagnostic methods have been developed to identify the aetiological agents of dermatophyte infections based on PCR (Luk et al 2012;Mehlig et al 2014), including PCR-restriction fragment length polymorphism assays (Rezaei-Matehkolaei et al 2012), reverse line blot hybridization assays (Zeng et al 2007;Bergmans et al 2008), oligonucleotide arrays (Han et al 2014), and real-time PCR (Arabatzis et al 2007;Bergman et al 2013;Kobylak et al 2015). Among these, PCR-based analysis has become the most reliable method for the identification of pathogenic fungi from clinical specimens (Kondori et al 2010).…”