Background: Foodborne illnesses continue to be a leading cause of morbidity and mortality worldwide; however, the burden of diseases caused by food-borne pathogens remains largely unknown. Objectives: The aim of the present study was to culture-confirmed the bacterial profile and their antibiotic resistant pattern in Food and Drug Laboratory, Alborz University of Medical Sciences, Karaj, Iran. Patients and Methods: A total of 22 bacteria including of Staphylococcus aureus, Klebsiella spp and E. coli were presumptive isolated from the traditional ice cream, cream pastries, sausage, and salami by the Official Food Microbiology Laboratory, Deputy of Food and Drug Administration, Alborz University of Medical Sciences, Karaj, Iran, and sent to the Research Center Laboratory, Alborz University of Medical Sciences, to confirm the bacterial spp by multiplex polymerase chain reaction. These isolates were also checked for their antimicrobial resistance pattern according to CLSI guideline. Results: The highest rate of contamination was with Klebsiella spp 09 (40.9%), followed by S. aureus 07 (31.8%), E. coli 06 (27.27%), as reported by the Official Food Microbiology Laboratory of Alborz University of Medical Sciences. Gel electrophoresis of the isolates shows the 600bp bp and 80 bp gene among S. aureus and E. coli respectively. The antibiotic resistant pattern in case of Klensiella spp showed that 6 (66.6%) Klensiella spp were resistant to Penicillin and Cotrimoxazole. Similarly, penicillin and amoxicillin were found the highest resistant antibiotic against 83.3% E. coli, however, ceftriaxone showed the highest sensitivity against 100% E. coli isolates. Conclusions: In conclusion, Klebsiella spp, S. aureus and E. coli are contaminants of food specimens obtained from food industries in Karaj, Iran; they constitute a serious health risk for human population. Moreover, the principal purpose of this study is to increase awareness of the antibiotic resistance of these bacteria poses threat.