Salmonellosis is one of the most important bacterial diseases affecting poultry. Its importance is derived from the loss in productivity in affected birds and the hazard it causes for public health. For preparation of polyvalent inactivated Salmonella vaccine, we tested the local virulent Salmonella Typhimurium, Salmonella Enteritidis and Salmonella Kentucky strains phenotypically by the microscope, biochemically by API test and genotypically by Polymerase Chain Reaction (PCR) using a specific primer for fliC gene of Salmonella Typhimurium and a specific primer for sefA gene of Salmonella Enteritidis and a specific primer for invA gene of Salmonella Kentucky. In Conclusion: Salmonellae are gram negative rods. They are aerobic or facultative anaerobic organisms that ferment sugars, producing gas and H2S and that have a complex antigenic structure (somatic "O", flagellar "H", and capsular "K" antigens). Multiplex PCR reveals that Salmonella Kentucky, Salmonella Typhimurium and Salmonella Enteritidis were positive for invA gene with a product of 284bp, fliC gene with a product of 620bp and sefA gene with a product of 488bp respectively.