Introduction: A nosocomial infection is “an infection acquired in the hospital by a patient who was admitted for a reason other than that infection” (WHO). It is necessary to prevent the infection in order to reduce its impact on morbidity and mortality in patients. The aim of this study is to assess the knowledge, attitude, and practice of health workers regarding nosocomial infection in selected hospitals in Birtamode Municipality, Jhapa.

Methods: This cross-sectional study was carried out among 198 individual health workers from selected hospitals in Birtamode Municipality through a multistage sampling technique. Knowledge, attitude, and practice of health workers regarding nosocomial infection were assessed with a validated, self-administered, pre-structured questionnaire. Statistical Package of Social Science version 23 was used to analyze the data, and the Chi-square test was applied to identify the association of dependent variables with independent variables.

Results: 198 health workers with a 100% response rate participated in the study. It was found that 76.8% had good knowledge, 75.3% had a positive attitude, and 97% had good practice regarding nosocomial infection transmission, prevention, and control during health care delivery in selected hospitals in Birtamode Municipality. There was a significant association between health workers’ knowledge and their working department, attitude and their working department, as well as working hours per week, practice, and their working department, with a p-value less than 0.05.

Conclusions: The knowledge, attitude, and practice of health workers regarding nosocomial infection were adequate while delivering health care services in selected hospitals in Birtamode Municipality.