title: Detection of common dermatophytes in clinical specimens using a simple quantitative real-time TaqMan polymerase chain reaction assay.
abstract: BACKGROUND
  Dermatophytes are common fungal pathogens causing mostly superficial infections in humans with a high prevalence worldwide. Traditional detection techniques are time-consuming and insensitive, whereas molecular detection methods have proved to be much more rapid and sensitive.
  
  
  OBJECTIVE
  To develop a modular singleplex quantitative real-time polymerase chain reaction (qRT-PCR) assay for the detection of the most common dermatophytes in clinical specimens.
  
  
  METHODS
  The qRT-PCR assay is based on single-tube reactions with TaqMan probes. We validated the test with 311 clinical samples of human and animal origin submitted for routine diagnosis and compared the qRT-PCR results with microscopy and culture.
  
  
  RESULTS
  qRT-PCR proved to be significantly more sensitive than microscopy and culture, with 21·2% more positive samples. Among the 201 dermatophytes identified 152 were Trichophyton rubrum (75·6%) and 34 were Trichophyton interdigitale (16·9%). Only 15 samples were determined as less common dermatophytes (Microsporum canis, Epidermophyton floccosum, Trichophyton verrucosum and Arthroderma benhamiae). In the present study, pathogen identification was achieved for 95·2% of all samples (including negatives) by applying only three detection tests (pan-dermatophyte, T. rubrum and T. interdigitale).
  
  
  CONCLUSIONS
  The qRT-PCR assay developed in this study allows the specific and sensitive detection of relevant dermatophytes at low cost in a short time.
authors.affiliation: National Reference Laboratory for Dermatophytes, Institute for Microbiology and Hygiene, Charité - Universitätsmedizin Berlin, Rahel-Hirsch-Weg 3, 10117, Berlin, Germany.
authors.forename: T
authors.initials: T
authors.lastname: Ohst
authors.affiliation: National Reference Laboratory for Dermatophytes, Institute for Microbiology and Hygiene, Charité - Universitätsmedizin Berlin, Rahel-Hirsch-Weg 3, 10117, Berlin, Germany.
authors.forename: C
authors.initials: C
authors.lastname: Kupsch
authors.affiliation: National Reference Laboratory for Dermatophytes, Institute for Microbiology and Hygiene, Charité - Universitätsmedizin Berlin, Rahel-Hirsch-Weg 3, 10117, Berlin, Germany.
authors.forename: Y
authors.initials: Y
authors.lastname: Gräser
mesh_terms: D000818:Animals; D003883:Arthrodermataceae / Q000235:genetics / Q000302:isolation & purification*; D004271:DNA, Fungal / Q000032:analysis; D003881:Dermatomycoses / Q000175:diagnosis*; D006197:Hair / Q000382:microbiology; D006201:Hair Diseases / Q000175:diagnosis; D006801:Humans; D009260:Nail Diseases / Q000175:diagnosis; D009262:Nails / Q000382:microbiology; D021141:Nucleic Acid Amplification Techniques; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D012867:Skin / Q000382:microbiology
chemical_list: D004271:DNA, Fungal