BackgroundPharmacogenomics is a field of science which studies the impact of inheritance on individual variation in medication therapy response.AimWe assessed healthcare professionals’ knowledge, attitude, and interest toward pharmacogenomics.MethodsA cross-sectional survey was conducted using a 32-item questionnaire among physicians, nurses, and pharmacists who were working at the University of Gondar Referral and Teaching Hospital in northwest Ethiopia. Descriptive statistics was applied, and the categorical variables were summarized as frequency and percentages. An analysis of variance (ANOVA) test was performed to compare mean scores among health professionals. A p-value of <0.05 was considered as statistically significant.ResultsOf 292 health professionals who responded, the majority were male (60%) and the mean age of study participants was 27.00 (±4.85 SD) years. The mean knowledge scores of all participants, pharmacists, physicians, and nurses were 2.343±1.109, 2.671±1.059, 2.375±1.093, and 2.173±1.110, respectively. Based on the ANOVA test, a statistically significant difference was noted in mean knowledge score between pharmacists and nurses (p=0.002). More than two-thirds (67.33%) of nurses, 42.86% of pharmacists, and 40.27% of physicians who participated did not know that genetic variations can account for as much as 95% of the variability in drug disposition and effects. The ability to accurately apply their knowledge to drug therapy selection, dosing, or monitoring parameter was reported by 35.3% of the participants. More than two-thirds (69.2%) of participants thought that pharmacogenomic testing will allow the identification of the right drug with less side effects. Most of the participants (83.2%) also requested to have training on pharmacogenomics.ConclusionParticipants showed limited knowledge, but they had positive attitude toward pharmacogenomics. Educational programs focusing on pharmacogenomic testing and its clinical application need to be emphasized.