Rapid and direct detection of Microsporum canis from canine hairs by PCR and PCR-RFLP

Abstract

Microsporum species are the most common agent of fungal dermatitis in dogs and cats and are transmitted to humans. The diagnosis of the disease is almost achieved by fungal culture and microscopic examination. But these methods take about two weeks to identify them. In this study, we amplified a part of the dermatophyte gene by PCR using TR, ITS, CHS1, dDP, MCN primer pairs. The result showed DNA concentration was increased depends on a number of hair. Each DNA sample was amplified with the primer sets corresponding to specific region of the Microsporum canis (TR; 580 bp, ITS; 720 bp, CHS1; 620 bp, dDP; 2380 bp, MCN; 639 bp) and restriction enzyme analysis (RFLP) of amplified products (TR, ITS, MCN) by SacI, SmaI, EcoRⅡ was done to identify species. The PCR products amplified from the DNA revealed each band corresponding to the respective amplified region of each fungus. All of 24 dermatophytosis patients were infected with Microsporum canis. The nucleotide sequencing on the PCR products (ITS, CHS) were performed and completely coincided with the reference sequences of the Microsporum canis. The PCR assay is quick and simply to directly detect Microsporum canis from the infected hair. This method is provide a sensitive and rapid detection and identification system for dermatophyte species, which may be applied to epidemiological surveys and routine practice.