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Background: Nosocomial infections remain a major global health concern, affecting an estimated 7% of hospitalised patients in developed countries and up to 10% in developing countries. Their consequences include prolonged hospital stays, increased antimicrobial resistance, elevated healthcare costs, and higher mortality. Although clinical staff such as radiographers, physicians, and nurses typically receive structured training in infection prevention, junior hospital staff are frequently overlooked in this regard, despite performing duties that place them in close and repeated contact with patients and contaminated materials. Few studies have specifically examined the knowledge and practices of junior staff with respect to nosocomial infection prevention and control. Objective: To assess the level of knowledge of nosocomial infections and associated prevention and control measures among junior staff of Federal Teaching Hospital Gombe. Methods: A descriptive cross-sectional census study was conducted among the population of junior staff (n = 80) across the hospital, comprising cleaners, porters, and health assistants. Data were collected using a structured, interviewer-administered questionnaire and analysed descriptively (frequencies and percentages) using SPSS version 26. Results: Only 6.3% (5 of 80) of respondents reported prior knowledge of nosocomial infections, although all respondents (100%) recognised the potential for infection transmission from patients and contaminated materials. Adherence to preventive practices was inconsistent: 46.3% reported always wearing gloves and face masks during cleaning, while 53.7% did so only occasionally. Reported barriers to consistent practice included limited access to training, heavy workloads, and inadequate budgetary allocation for infection prevention materials. Conclusion: The study demonstrated a low level of knowledge and inconsistent application of nosocomial infection prevention and control measures among the junior hospital staff. Despite recognising their role in infection transmission, gaps in training, workload challenges, and limited access to preventive resources affected compliance with recommended practices. Regular, structured infection prevention training and improved institutional support are essential to strengthen infection prevention and control practices and enhance patient and staff safety.