PCR-RFLP: an alternative to culture (gold standard) in diagnosis of dermatophytes in dogs and cats

Abstract

ABSTRACT. Dermatophytes are fungi keratinophilic and keratinolytic that causing dermatophytosis in humans and animal and, even with a variety of available methods, the laboratory diagnosis is still a difficulty in the veterinary clinic routine. The aim of this study was to evaluate the PCR-RFLP technique as an alternative method to the culture (gold standard) for the diagnosis of dermatophytes in dogs and cats. Were analyzed 150 samples from animals with skin disease. Culture and DNA extraction from fur and/or crusts were performed. To the PCR-RLFP positive controls were used colonies of M. canis (URM 6273), M. gypseum (URM 6921) and T. mentagrophytes (URM 6211), from the URM Micoteca - Department of Mycology, Biological Sciences Center of the Universidade Federal do Pernambuco. The primers used were ITS1 and ITS 4. The BseBI endonuclease was selected to obtain fragments pattern. The Kappa coefficient (K) was used to analyze the agreement between culture and PCR-RFLP. There was an almost perfect agreement (K = 0.813) between the results of culture and PCR-RFLP. The molecular technique showed an interesting potential to the dermatophytes identification in clinical samples from dogs and cats. However, further studies are required to guide the future of PCR-RFLP in the laboratory routine.