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Introduction: Typhoid fever (TF) is a febrile global health problem caused by Salmonella enterica serovar Typhi ( S. Typhi) with relatively high prevalence in low- and middle-income countries including Ethiopia. Identifying local prevalence and gaps in knowledge, attitude, and practice (KAP) towards TF is recommended by the World Health Organization to implement preventive measures. Therefore, this study determined the prevalence of S . Typhi and KAP of febrile patients towards TF in Injibara General Hospital, Northwest Ethiopia., Methods: Hospital-based cross-sectional study was conducted from January to March 2020. A total of 237 patients were included conveniently. Data on KAP and demographic variables were collected using a structured questionnaire by face-to-face interview. After the interview, 5 ml venous blood was collected and processed using the Widal test following the manufacturer's instruction. Mean scores and percentages were used to determine the level of KAP. Multivariable analysis was done to correlate KAPs with TF. P value < 0.05 was considered statistically significant., Results: The overall prevalence of S . Typhi was 25.7%. The highest seroprevalence was observed among the age group of 30-34 years (33.3%) and patients with no education. The majority of participants know the major ways of TF transmission (59.1-90.7%) and prevention (81.4%) methods. However, the misconception on the route of TF transmission was observed in 13.5-36.7% of participants. About 65.4% and 67.5% of study participants were considered knowledgeable and had good preventive practice towards TF, respectively. Being a student (AOR = 0.227, CI = 0.053 - 0.965) and considering mosquito bite as transmission routes (AOR = 2.618, CI = 1.097 - 6.248) were significantly associated with TF., Conclusion: High S . Typhi prevalence was observed in the study area. Moreover, the misconception on the transmission of typhoid fever and educational level was a risk factor for TF. Thus, health facilities should incorporate topics on typhoid fever as part of their health education system within health facilities and in the community., Competing Interests: The authors declare that they have no conflict of interest., (Copyright © 2021 Wondemagegn Mulu et al.)