Direct detection of five common dermatophyte species in clinical samples using a rapid and sensitive 24-h PCR-ELISA technique open to protocol transfer

Barbara Beifuss, Guntram Bezold, Petra Gottlöber, Claudia Borelli, Jeanette Wagener, Martin Schaller, Hans Christian Korting

Research output: Contribution to journalArticlepeer-review

47 Citations (Scopus)

Abstract

Identification of dermatophytes is usually based on morphological characteristics determined by time-consuming microscopic and cultural examinations. An effective PCR-ELISA method has been developed for rapid detection of dermatophyte species directly from clinical specimens within 24 h. Isolated genomic DNA of skin scrapings and nail samples from patients with suspected dermatophyte infections is amplified with species-specific digoxigenin-labelled primers targeting the topoisomerase II gene. The subsequent ELISA procedure with biotin-labelled probes allows a sensitive and specific identification of the five common dermatophytes -Trichophyton rubrum, T. interdigitale, T. violaceum, Microsporum canis and Epidermophyton floccosum. PCR-ELISA, based on the new polyphasic species concept, was assessed using 204 microscopy-positive samples in two university mycological laboratories in Munich and Tübingen, and 316 consecutive specimens - regardless of mycological findings - in a dermatological practice laboratory in Neu-Ulm. One of the five dermatophytes was confirmed by PCR-ELISA in 163 of 204 (79.9%) of the clinical samples from the university hospitals found positive using microscopy. Culture was positive for dermatophytes in 59.8% of the same cases. A significant difference between these two methods could be demonstrated using the McNemar test (P <0.005). Analysis of specimens from Neu-Ulm confirmed the results in a dermatological practice laboratory as 25.0% of the specimens had positive PCR results, whereas only 7.3% were positive according to culture. Direct DNA isolation from clinical specimens and the PCR-ELISA method employed in this study provide a rapid, reproducible and sensitive tool for detection and discrimination of five major dermatophytes at species level, independent of morphological and biochemical characteristics.
Original languageEnglish
Pages (from-to)137-145
Number of pages9
JournalMycoses
Volume54
Issue number2
Early online date27 Sep 2009
DOIs
Publication statusPublished - Mar 2011

Keywords

  • Arthrodermataceae
  • DNA Primers
  • DNA, Fungal
  • Dermatomycoses
  • Enzyme-Linked Immunosorbent Assay
  • Humans
  • Molecular Sequence Data
  • Polymerase Chain Reaction
  • Sensitivity and Specificity

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