During 2010-2011, Real-time PCR procedure was used to detecting FMDV RNA on 147 epithelium samples from the field. In this survey, for Real-time PCR from 3D gene segment as conserve region selected for tracking all of seven serotypes FMDV. The assay detected the viral RNA in all serotypes of FMDV. The rRT-PCR specifically detected FMD virus in sample with greater sensitivity than our conventional RT-PCR procedure and our routine diagnostic procedures. Sensitivity of this technique was estimated by 10-log serials dilution Asia virus RNA that defined 101.2 TCID50/ml. Also, to assess the specificity of primer pairs related to 3D segment and specific probe was used Swine vesicular disease virus (SVDV) and bluetongue virus (BTV) isolates RNA. The results showed that rRT-PCR is seen as a powerful and valuable tool concomitant with routine diagnostic methods for FMD virus diagnosis.