Objectives: This study aimed to assess malaria seroprevalence among acute febrile illness cases who come for health care seeking in the high malaria-endemic setting of North West Ethiopia. Methods: Institutional-based descriptive serosurvey of malaria infections was employed among 18,386 febrile patients from September 2020 to August 2021. Data were entered using Epi Data version 4.2 and exported to STATA (SE) R-14 version statistical software for further analysis. Bi-variable and multivariable regression analyses were conducted to identify malaria infection. Finally, variables with P-value less than 0.05 were considered significant predictors for malaria infection. Results: The mean (±standard deviation) age of participants was 48.6 (±18.4) years. The overall seroprevalence of malaria infection was estimated as 27.8% (95% confidence interval = 27.2; 28.6, standard error = 0.003). Malaria infection was significantly associated with participants being female (adjusted odds ratio = 2.9; 95% confidence interval = 1.8; 3.7, P = 0.01), age 5–29 years (adjusted odds ratio = 2.2; 95% confidence interval = 1.7; 2.8, P = 0.02), rural (adjusted odds ratio = 3.9; 95% confidence interval = 1.9; 4.4, P = 0.001), and Hgb ⩽11 mg/dL (adjusted odds ratio = 3.4; 95% confidence interval = 1.9; 5.86, P = 0.01). Conclusion: Nearly every three to ten acute febrile cases were positive for confirmed malaria infection. The risk of malaria infection was significantly associated with respondents being female, aged 5–29 years, rural, and levels of hemoglobin were significantly associated with malaria infection.