Background: Salmonellosis is increasingly recognized as a worldwide public health concern. Salmonella Infantis can infect both humans and animals, including poultry. It has been one of the most reported isolated serovars from different parts of the world. Although some research has been carried out on the pathogenesis of S. Infantis, little scientific understanding of its pathogenesis is available.Objectives: This study aimed to analyze the virulence genes of S. Infantis recovered from different sources of poultry in Iran.Methods: Six virulence genes of 54 S. Infantis strains originated from broiler feces, poultry processing, and broiler carcasses were examined. Gene-specific polymerase chain reactions were designed and employed to detect the presence or absence of 6 important virulence genes (sopB, sopE, sitC, pefA, sipA, and spvC) in 54 S. Infantis isolates.Results: In this study, sopE, sitC, pefA, sipA, and sopB virulence genes were detected in 51(94.4%), 49(90.7%), 26(48.1%), 15(27.7%), and 5(9.2%) isolates, respectively. The spvC gene was not detected in any of the isolates. Conclusion: In the present study, a remarkably identical profile was found on virulence genes’ presence in isolates recovered from broiler feces and poultry processing plant sources, that is a public health concern. However, more S. Infantis isolates from various poultry sources, and human origin should be examined and analyzed. The findings of this survey can help the health researchers better understand the pathogenesis and epidemiology of S. Infantis in Iran.