Evaluation of a PCR melting profile method for intraspecies differentiation of Trichophyton rubrum and Trichophyton interdigitale.
In order to identify the source of infections caused by dermatophytes, as well as the pathogen transmission pathway, there is a need to determine methods that allow detailed genetic differentiation of the strains within the dermatophyte genera. In this work, a PCR melting profile (PCR-MP) technique based on the ligation of adaptors and the difference in melting temperatures of DNA restriction fragments was used for the first time for intraspecies genotyping of dermatophytes. Clinical isolates and reference strains of dermatophytes isolated from skin, scalp, toenails and fingernails were used for this study. PCR-MP and random amplification of polymorphic DNA (RAPD) were used to type 11 isolates of Trichophyton rubrum, 40 isolates of Trichophyton interdigitale and 14 isolates of Microsporum canis. The results distinguished five types (containing one subtype) characteristic for T. rubrum and seven types characteristic for T. interdigitale using the PCR-MP technique. Analysis conducted using RAPD revealed five types for T. rubrum and four types for T. interdigitale isolates. No differentiation was observed for the M. canis isolates with either method. These results demonstrate that PCR-MP is a reliable method for the differentiation of T. rubrum and T. interdigitale strains and yields a discriminatory power that is at least equal to that of RAPD.
Leibner-Ciszak
Justyna
J
Gdansk University of Technology, Chemical Faculty, Department of Microbiology, ul. Narutowicza 11/12, 80-233 Gdansk, Poland.
Dobrowolska
Anita
A
University of Lodz, Department of Genetics of Microorganisms, ul. Banacha 12/16, 90-237 Lodz, Poland.
Krawczyk
Beata
B
Gdansk University of Technology, Chemical Faculty, Department of Microbiology, ul. Narutowicza 11/12, 80-233 Gdansk, Poland.
Kaszuba
Aleksandra
A
Medical University of Lodz, Department of Dermatology and Venerology, ul. Kniaziewicza 1/5, 91-347 Lodz, Poland.
Stączek
Paweł
P
University of Lodz, Department of Genetics of Microorganisms, ul. Banacha 12/16, 90-237 Lodz, Poland.
D004271:DNA, Fungal / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D008864:Microsporum / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D009691:Nucleic Acid Denaturation; D016133:Polymerase Chain Reaction / Q000379:methods*; D013045:Species Specificity; D014005:Tinea / Q000382:microbiology; D014249:Trichophyton / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification
D004271:DNA, Fungal