To examine the knowledge and practice, as well as factors influencing universal precautions practices amongst Nigerian House officers and Nurses. A Cross-sectional descriptive study. Sample selection was by stratified random sampling. Information was elicited using pretested, structured, self-administered questionnaire. Data were analyzed using EPI-info. Most Doctors, 66.6%, were aged 26-30 years while the Nurses, 41.1%, were aged 40 years. 57.6% of the Doctors were males, while 85.7% of the nurses were females. Knowledge of universal precautions measures was high for both categories of respondents--97.0% for doctors and 92.0% for nurses, although practice was better for the nurses, 75.0%, compared to the doctors, 15.2%, p < 0.05. The most important factor influencing universal precautions practice is the lack of provision of adequate protective equipments. Other factors, all of which show significant difference between the doctors and nurses (p < 0.05), include carelessness; lack of display of universal precautions guidelines; emergency nature of the procedure; insufficient water supply; patient perceived to be at low risk of blood borne pathogens; pressure of time; and universal precautions equipments interfering with technical skills. Although knowledge of universal precautions is high for both house officers and nurses, practice is however better amongst the latter than the former. The effective knowledge and practice of universal precautions amongst hospital workers are of absolute necessity to prevent infections from blood and body fluid pathogens.