Nosocomial infections remain a significant public health concern, making prevention among healthcare workers essential to reduce associated risks. This study investigated the factors that influence preventive practices against hospital-acquired infections among healthcare personnel in the Rivers East Senatorial District of Rivers State. A descriptive cross-sectional design was adopted, targeting a population of 2,078 primary healthcare workers. Using a multistage sampling technique, 1,142 participants were selected for the study. Data were gathered with a researcher-developed instrument known as the “Nosocomial Infection Prevention Questionnaire (NIPQ),” which had a reliability coefficient of 0.78. Statistical analysis was conducted using SPSS version 27.0, and both research questions and hypotheses were examined through Linear Regression at a 0.05 significance level. Findings revealed that self-efficacy accounted for 61.4% of the variance in preventive behaviour (R² = 0.61, r = 0.75); cues to action explained 70.3% (R² = 0.70, r = 0.75); and perceived barriers contributed 58.9% (R² = 0.58, r = 0.68) to nosocomial infection prevention among healthcare workers in the district. Based on various analyses of data, nosocomial infection prevention among healthcare workers in Rivers East Senatorial District was predicted by several factors,, with the most profound being self-efficacy and perceived barriers. It was recommended among others that the primary healthcare management agency should come up with an assessment method to ensure adequate compliance with nosocomial infection prevention, with the view of eliminating every barrier identified.