The prevalence of bacterial presences on the handles of car doors in Al-Najaf Province was evaluated. The current study was carried out between December 2017 and March 2018. A total of 100 samples were collected and cultured for bacterial isolation. The incidence of positive specimens was as the follow; (40%) with bacterial cultures, (52%) with yeasts cultures and (8%) with negative aerobic cultures. In addition, the current results involved that (70%) of isolates was Gram positive and (30%) of isolates was Gram negative. The most isolated bacteria were S. aureus, Klebsiella sp., E. coli, Proteus sp., Salmonella sp., Bacillus sp., and Streptococcus sp.  The current study was indicated high prevalence rate of bacteria on the handles of car doors in Al-Najaf Province.