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Background: Gastroenteritis due to Salmonella is common in human and considered as a global ‎dilemma of public health. This study was done to determine cmlA/tetR, ‎‌bla PSE-1, bla TEM and sip B ‎in the Salmonella strains by Multiplex-PCR method and their antibiotic susceptibility profile. ‎ Materials and methods: In this cross-sectional study, 163 clinical samples were obtained from ‎patients admitted to Hazrat-e Rasool General Hospital. The antibiotic susceptibility test was determined ‎using the disk diffusion method agreeing with CLSI guideline. Then, M-PCR was achieved for ‎determination of these target genes by the specific oligonucleotides primers. ‎ Results: Of 163 collected samples, 48(29.4%) Salmonella spp., were obtained, which 25(52.1%) were ‎S. enteritidis, 14(29.2%) S. typhimurium and 9(18.7%) S. infantis. Antibiotic resistance analysis showed ‎that the highest resistance rate were related to tetracycline (n: 27, 56.2%) and then streptomycin and ‎chloramphenicol (n: 15, 31.2%). All isolates (n: 48, 100%) were susceptible to imipenem, amikacin, ‎gentamycin and trimethoprim/sulfamethoxazole. The MPCR results revealed that 62.5% and 16.6% of ‎Salmonella spp., isolates carried cml/tetR and sipB genes, respectively. ‎ Conclusion: According to our results, detection and genotyping of virulence genes and comparison ‎with global ranging is a basic requirement in the control and prevention of salmonellosis in industrial ‎purposes.‎ Keywords: cmlA/tetR, PSE-1, TEM, sip B, Salmonella, Multiplex-PCR