BACKGROUND: Yersinia ruckeri is the etiological agent of enteric red mouth (ERM) or yersinioisis disease, one of the important bacterial diseases in the cultured salmonids. OBJECTIVES: The purpose of present study was detection of gyrA gene (quinolone resistance) in the Y. ruckeri bacterium. METHODS: In this study fish were evaluated in average size 8-12 cm from six rainbow trout farms in Chahar Mahal va Bakhtiyari province (Iran). In each farm 10 fish (totally 60) suspected to yersinioisis were randomly selected; sampling was done from lower part of intestine and cultured on Trpticase Soy Agar (TSA). The mediums were transferred to incubator and kept at 22 °C for 48 hours. Pure colonies which are grown on the mediums were tested by catalase, oxidase and gram staining, then those of gram-negative, catalase positive and  oxidase negative were diagnosed, and cultured on Waltman- Shots medium (as specific medium for Y. ruckeri). These mediums were incubated at 22 °C for 48 h. Colonies that were grown were tested by PCR method for Y.ruckeri detection. Then, in the identified strains of Y.ruckeri gyrA gene were detected by PCR test. RESULTS: The results of bacteriological, biochemical and molecular tests showed that three cases out of total isolates were identified as Y. ruckeri. In all isolates of Y. ruckeri, gyrA gene was identified by molecular test. CONCLUSIONS: Identification of quinolone resistance gene in Y. ruckeri isolates can be the reason of low efficacy of these classes of antibiotics in the aquaculture. ِTherefore, the policy of treatment should be changed specially in enteric red mouth disease.