Introduction: Hospital Acquired Infections are a major global safety concern for patients, healthcarernprofessionals and the community. Health care workers had potentially exposed to infectious materialsrnsuch as blood, tissue, specific body fluids, medical supplies, equipment or environmental surfacesrncontaminated with these substances.rnObjective: The objective of this study was to assess infection prevention practices among healthrnprofessionals and associated factors in Tikur Anbessa Specialized Hospital, 2019.rnMethods: A cross-sectional study was conducted at Tikur Anbessa Specialized Hospital from Marchrnto April 2019. The study populations were Health care professionals selected using simple randomrnsampling. Data was collected using self-administered questionnaire supplemented by observation. Datarnwas entered in to Epi version 7.2.1 and exported to SPSS version 22 for further analysis. Logisticrnregression analysis was used to identify factors associated to with infection prevention practices.rnFindings were presented using odds ratio with 95% confidence intervals. A P-value less than 0.05 wasrnused to declare statistical significance. rnResults: 396(94.5%) Health care professionals participated in the study. The mean age of participantsrnwas 28.54 years, and 188 (47.5%) were males and 208(52.5%) were females. 305(77%) of healthrncare professionals are knowledgeable and 281(71%) of the respondents were found to be positivernattitude towards infection prevention. 230(53.8%, (95%CI: 49.0-58.1)) had good infection preventionrnpractices. Having good knowledge on infection prevention measures (AOR =3.66,rn95%CI:(2.005,6.704)), having infection prevention guidelines on infection prevention practicesrn(AOR=1.926, 95%CI:(1.148,3.230)) and average working hours per week (AOR=1.866, 95%CI:rn(1.037,3.356)) were predictors of infection prevention practices.rnConclusion: The results of the study show that half of (53.8%) HCPs had good infection preventionrnpractices. Having good knowledge, availability of infection prevention guideline and average workingrnhour per week of health professionals are predictors of good infection prevention practices. As a result,rnto sustain and improve good infection prevention practices, adequate pre-service as well as on jobrntrainings should be given and accessing infection prevention guidelines in each department.