Literatures proved that Hand hygiene is the most important and effective infection prevention and control measure to prevent the spread of microorganisms causing HAIs and improving hand hygiene is consider a vital intervention to promote optimum patient safety in delivery room. Aim of the study: This study conducted to assess hand hygiene practices of health care personnel in the delivery room at the middle Euphrates teaching hospitals. Methods: A Descriptive qusi-expremental research design begin in 20th February to 26th May 2016, Current study sample involve all midwives and physicians in the delivery room (Total coverage.). Questionnaire used for data collection by interview forms and observational checklist was obtain from the extensive review of relevant literature and related studies Data analyzed through utilize (SPSS) software version (16) where, included descriptive analysis and inferential data analysis. The study conducted among 37 physicians and 97 midwives working in the delivery room are females. The current study indicate that the overall evaluation for the health staff practices regarding hand hygiene is fair at Karbala, Al-Najaf, Babylon and Diwaniah with high difference in health staff practices regarding infection control precautions (hand hygiene) and the different studied governorates at p-value 0.001, based on the finding of present study majority of health care personnel have fair applies related to hand hygiene practices at different studied governorate hospitals. Updating practice of health care personnel through continuing in-service educational programs. Regular inspection and follow-up from the ministry of health for assurances good hand hygiene, the important of exist motivation system and punishment system to the neglected health care personnel.