Introduction: The one of the main causes of hospital acquired infection (HAI) is the lack of knowledge and improper practice of employees in the hospital. Training programs on the infection control for each employees can reduce the rate of infection acquired from hospital. Aim of the study: The aim of this study was to assess the knowledge and practice of infection control among the employees in DMCH and ShSMCH. Methods: This descriptive type of cross-sectional study was conducted in Dhaka Medical college Hospital and Shaheed suhrawardi Medical college Hospital Dhaka for a period of one year starting from January-2019 to December-2019. Total 139 employees (Physician and Nurses) were included in this study. Result: Among 139 respondents, mean age were 31.97±7.412 (SD). 51.8% (n= 72) were female and 48.2% (n= 67) were male. In this study, 99.3% (n=138) of the respondents were aware of infection control. The current study shows that the 100% (n=139) of respondents know about nosocomial infection. 95.7% (n=114) know infection control technique prevent nosocomial infection and 82% (n=114) of the respondents know infection can occur after discharge from hospital. Maximum 63.3% (n=88) of the respondents know the most effective method to control infection were hand washing properly and the minimum 4.3% (n=6) know prudent use of antibiotics, 38,8% (n=54) know wearing caps, masks and shoe cover and 9.4% (n=13) knows regular vaccination of health care worker. Maximum of the respondents 48.2% (n=67) practice often, 33.8% (n=47) practice always, 17.3% (n=24) practice sometime and minimum .7% (n=1) of the respondent never practice infection control guideline. Conclusion: Physician and nurses in the current study have average knowledge and practice level regarding infection control.