Evaluation of a PCR melting profile method for intraspecies differentiation of Trichophyton rubrum and Trichophyton interdigitale

• DOI: 10.1099/jmm.0.013458-0
• Publication Date: 2009-11-06T03:04:21Z
• Authors: Justyna Leibner-C...; Anita Dobrowolska; Beata Krawczyk; Aleksandra Kaszuba; Paweł Stączek
• Keywords: 0303 health sciences; 03 medical and health sciences; Species Specificity; Tinea; Trichophyton; Humans; Microsporum; DNA, Fungal; Nucleic Acid Denaturation; Polymerase Chain Reaction

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 ofTrichophyton rubrum, 40 isolates ofTrichophyton interdigitaleand 14 isolates ofMicrosporum canis. The results distinguished five types (containing one subtype) characteristic forT. rubrumand seven types characteristic forT. interdigitaleusing the PCR-MP technique. Analysis conducted using RAPD revealed five types forT. rubrumand four types forT. interdigitaleisolates. No differentiation was observed for theM. canisisolates with either method. These results demonstrate that PCR-MP is a reliable method for the differentiation ofT. rubrumandT. interdigitalestrains and yields a discriminatory power that is at least equal to that of RAPD.