Salmonella enterica serovar Gallinarum is the causative agent of fowl typhoid (FT) and Salmonella enterica serovar Pullorum is pullorum disease (PD), a severe systemic disease of chick and it has the same antigenic fomula, the close relation but distinct pathogen. The traditional bacteriologic and serologic methods routinely used but tedious, time consuming. some of biochemical differences are helpful in differentiating the two organisms, however variation in the characteristics of some strains can be observed. During 2006 to 2008, there was isolated 30 strains. The biochemical characteristics of S. Gallinarum was nonmotile, fermentation of dulcitol, maltose but positive arginine (6.6%), lysine (83.3%) and arabinose (20.0%). The antimicrobial susceptibility test showed 100% sensitive to amikacin, ampicillin, amoxicillin/clavulanic acid and florfenicol, but resistant to penicillin (100%) and erythromycin (60.0%). This PCR method can be applied in the diagnosis between S. Gallinarum and S. Pullorum.