TY - JOUR
T1 - Rapid identification of Trichophyton tonsurans by PCR-RFLP analysis of ribosomal DNA regions
AU - Mochizuki, Takashi
AU - Tanabe, Hiroshi
AU - Kawasaki, Masako
AU - Jackson, Colin J.
AU - Ishizaki, Hiroshi
N1 - Mochizuki, T., Ranabe, H., Kawasaki, M., Ishizaki, H., Jackson, C. J. (2003). Rapid identification of Trichophyton tonsurans by PCR-RFLP analysis of ribosomal DNA regions. Journal of Dermatological Science, Elsevier, 32, (1), 25-32
PY - 2003/6/1
Y1 - 2003/6/1
N2 - 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.
AB - 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.
KW - PCR-RFLP analysis
KW - indentification
KW - dermatophytes
KW - ribosomal DNA
KW - restriction enzyme analysis
KW - Trichophyton tonsurans
U2 - 10.1016/S0923-1811(03)00030-6
DO - 10.1016/S0923-1811(03)00030-6
M3 - Article
SN - 0923-1811
VL - 32
SP - 25
EP - 32
JO - Journal of Dermatological Science
JF - Journal of Dermatological Science
IS - 1
ER -