Identification of common species of dermatophytes by PCR-RFLP

To establish a simple, sensitive and effective technique for the identification of six common dermatophytes, polymerase chain reaction (PCR) and PCR0restriction fragment length polymorphism (RFLP) targeting Topoisomerase II gene were used. The DNA of 6 common dermatophytes was amplified by primer dPsD1 and then primers dPsD2. The products generated by dPsD2 were digested with restrictiion enzymeHinc II andHinf I separately. A DNA fragment of about 3390 bp was amplified by using primer dPsD1 from the genomic DNA of each dermatophyte species. The product of dPsD2 was 2380 bp and the restriction profiles ofHinc II andHinf I were between 58–1670 bp. By using PCR-RFLP, all of the 6 dermatophytoses were diagnosed to species level and no obvious difference identification betweenHinc II andHinf I. It is concluded that the PCR-RFLP identification of dermatophytes byHinc II orHinf I is efficient and rapid in clinical practice.