Rapid identification of Trichophyton tonsurans by PCR-RFLP analysis of ribosomal DNA regions

Takashi Mochizuki, Hiroshi Tanabe, Masako Kawasaki, Colin J. Jackson, Hiroshi Ishizaki

Research output: Contribution to journalArticlepeer-review

72 Citations (SciVal)

Abstract

Background: Culture morphology of Trichophyton (T.) tonsurans, an emerging pathogen of dermatophytosis in Japan, varies widely and species level identification is sometimes very difficult. Reliable molecular markers are expected to be introduced for their identification. Objective: The present study was conducted to evaluate the efficacy of restriction fragment length polymorphism (RFLP) analysis of PCR amplified ribosomal (r) DNA including internal transcribed spacers (ITS), as an identification tool. Methods: Total cellular DNA was extracted from 26 Japanese isolates of T. tonsurans, along with several taxa of the members in the T. mentagrophytes complex, T. rubrum, T. violaceum and Epidermophyton floccosum, using a mini-preparation method. PCR amplicons were digested with restriction enzymes Mva I or Hinf I, then electrophoresed on 5% polyacrylamide gel. Results: The banding profiles were observed about 8 h from initiating DNA extraction. Intraspecies polymorphism was not detected among T. tonsurans isolates, and their profiles obtained using Mva I digestion were clearly different from those of the other dermatophyte species. The restriction profiles evaluated from nucleotide sequence of the regions by a computer analysis were compatible with the electrophoresed profiles on gel. Conclusion: PCR-RFLP analysis is a rapid and reliable tool for the identification of T. tonsurans.
Original languageEnglish
Pages (from-to)25-32
Number of pages8
JournalJournal of Dermatological Science
Volume32
Issue number1
Early online date27 Feb 2003
DOIs
Publication statusPublished - 01 Jun 2003

Keywords

  • PCR-RFLP analysis
  • indentification
  • dermatophytes
  • ribosomal DNA
  • restriction enzyme analysis
  • Trichophyton tonsurans

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