The current research focused on the detection of Salmonella typhi and its relationship with the formation of gallstone and gall bladder cancer. Samples were collected from patients aged between 32-67 years (males and females) in Mosul city hospitals. The samples included 30gallbladder fresh tissues from patients suffering from gallstone and 20 formalinfixed-paraffin embedded (FFPE) gallbladder tissue from patients confirmed with gallbladder cancer. The results showed that 33% S. typhi isolates were diagnosed from the tissue samples using conventional methods, biochemical tests and Vitek2. All fresh tissues samples gave positive PCR results for the presence of FliC-d and CdtB genes and 46% positivity for S. typhi compared with conventional methods, 64% for females, 35% for males and 40 % positive in the gallbladder cancer of which 62% were females and 37%males.