Summary:Staphylococcus aureus and S. intermedius are considered as the most significant species causing skin infections in dogs. The aim of this study was to develop a Polymerase Chain Reaction technique for the detection of siet (S. intermedius exfoliative toxin) gene encoding exfoliative toxin in S. intermedius and to investigate its presence in S. intermedius isolates from dogs with pyoderma. A total of 41 isolates (35 S. intermedius, 4 S. aureus, one S. capitis subsp. ureolytica and one S. chromogenes) from dogs with pyoderma were included in the study. Original primers specifically amplifying 145 bp of siet gene and 182 bp of agr (accessory gene regulator) gene locus of S. intermedius were designed in the study. agr gene was detected in all S. intermedius isolates, but not in other isolates. siet gene was detected in all S. intermedius isolates. siet Polymerase Chain Reaction assay was found to be specific since no amplifications were observed with siet negative S. intermedius and other bacterial control strains. Rapid and reliable detection of staphylococci causing skin lesions in dogs and their virulence markers like siet gene will provide important data for clinical practice to manage the disease more effectively by means of treatment and prevention.Key words: Canine pyoderma, exfoliative toxin, siet gene, Staphylococcus intermedius. Piyodermalı köpeklerden izole edilen Staphylococcus intermedius'larda eksfoliatif toksin varlığının moleküler olarak belirlenmesi