Typhoid is a major public health concern. Even though antibiotics are usually used to treat typhoid fever, the spread of multi drug resistant Salmonella typhi is making antibiotics much less effective. This study was conducted to assess the prevalence of multidrug-resistant Salmonella typhi from the clinical samples. During this study, 154 blood samples of suspected typhoid patients were collected from the hospital and diagnostic center located in Chattogram City, Bangladesh. Isolation and identification of Salmonella typhi was done by both biochemical tests. PCR analysis was also done for the confirmation of biochemical result. Antimicrobial susceptibility test was performed according to the Kirby-Bauer disk diffusion method against ampicillin, chloramphenicol, cefepime, cotrimoxazole, ceptriaxone, ciprofloxacin, nalidixic acid, and azithtomycin. Out of 154, 21 (13.64%) isolates were identified as Salmonella typhi and the prevalence of typhoid in Chattogram, Bangladesh was 13.64% (n = 21). It was also found that children under the age of 5 are the more vulnerable target of Salmonella typhi infection. Antibiotic resistance profiling revealed 85% isolates were Multi-Drug Resistant (MDR) and highest resistance was found in case of Nalidixic acid. Although, most of the isolated Salmonella typhi were MDR, first generation antibiotics Co-trimoxazile, Chloramphenicol, and Ampicillin were found effective against Salmonella typhi.