Background:The prevalence of typhoid fever in Cameroon is high. However, studies on susceptibility of Salmonella to the recommended antibiotics are few and not extensive. This study investigated the susceptibility of Salmonella isolated from clinical specimens in the South West region of Cameroon and the genes responsible for their antibiotic resistance. Methods: Salmonellawas isolated from clinical specimens collected from some medical laboratories in the study area by culture on Salmonella Shigella agar. Identification and resistance genes analysis were done using biochemical tests, PCR and gene sequencing. Disc diffusion test was done using 11 commercial antibiotics and minimum inhibitory concentrations determined by microdilution test to assess antibiotic susceptibility profile. The Multi-Antibiotic Resistance (MAR) index values were calculated. Results: Salmonella was isolated from 50 out of 115 specimens, of which 46 (92 %) were multidrug resistant (MDR) with MAR indices > 0.2.The MDR isolates showed high resistance to penicillins and low resistance to ciprofloxacin. PCR revealed that antibiotic resistance was largely due to the tem-1, Sul 1, dfrA1, floR genes. Sequencing revealed double mutations at codons 83 (Ser83-Phe) and 87 (Asp87-Gly) and one mutation at codon 83 (Ser83-Trp) in the quinolone resistance determining regions of the gyrA gene in two isolates. Conclusion: This study has revealed presence of MDR Salmonella enterica isolates in Southwest Cameroon with several resistance genes. This finding will guide use of antibiotics in treatment of Salmonella infections in the study area and justifies the search for new efficacious antibacterials against the MDR strains.