BackgroundFailure to follow proper infection prevention practices puts healthcare workers, patients and communities at risk. Despite the increases of highly contagious infections, infection prevention practices among healthcare workers is unknown in many developing countries. The need to understand infection prevention practices is important for prevention and control of nosocomial infections. Therefore, the objective of this study was to assess infection prevention practices and associated factors among healthcare workers in governmental healthcare facilities in Addis Ababa, Ethiopia.MethodsA facility based cross-sectional study design was conducted from February to March 2016, in Addis Ababa among 629 healthcare workers who were selected by multistage sampling technique from 30 governmental healthcare facilities. Data were collected using pre-tested interviewer administered structured questionnaire. Data were entered into Epi -data 3.1 and exported to SPSS version 20 for analysis. Multivariable logistic regression model was used to identify factors associated with infection prevention practices.. Findings were presented using odds ratios with their 95% confidence intervals., A p-value less than 0.05 were used to declare statistical significance.ResultsSix hundred five (96.2%) healthcare workers participated in the study. Four hundred (66.1%, (95%CI: 62.1%–70.1%)) health care workers had good infection prevention practices. Having good knowledge on infection prevention meausures (AOR =1.53, 95%CI: 1.05–2.22), having positive attitude towards infection prevention practices (AOR=2.03, 95%CI: 1.26–3.26), having awareness on availability of standard operating procedures (AOR=1.97, 95%CI: 1.34–2.93) and presence of continuous water supply (AOR=1.68, 95%CI: 1.11–2.56) were predictors of good infection prevention practices.ConclusionTwo-third of the healthcare workers had good infection prevention practices. Having good knowledge on infection prevention measures, having positive attitude towards infection prevention practices, having awareness on availability of standard operating procedures and presence of continuous water supply were predictors of good infection prevention practices. To sustain good practices, adequate pre-service and in-service training should be in place to equip and update health care workers about infection prevention precautions. The need for continuous supervision should be implemented to strengthen adherence for infection prevention practices among workers along with sustainable and reliable water supply is crucial.