Standard precautions (SP) are essential in preventing transmission of infectious agents in hospital settings. This study is aimed at identifying perceptions of nosocomial infections, compliance with SP and associated factors among healthcare workers (HCWs). It was cross-sectional study done in 2014 at University of Nigeria Teaching Hospital. A pre-tested questionnaire was used. Out of 629, most were females (64.4%), nurses (46.1%) and within 20 – 59 years age range. One hundred and twelve (17.8%) have had at least one nosocomial infection since employment. Although many received training on SP (62%), only 20.3% were aware of hospital policy that enhances compliance. Regular access to PPEs (56.1%) and hand hygiene (62.8%) were reported. Most (64.5%) reported total absence of control measures for respiratory infections. Presence of an epidemic, hospital policies and managing an infectious patient encourage compliance with SP while emergency situations and non-availability of PPEs were major constriants. Nosocomial infections occurred most among orderlies (33.3%) and least among doctors (14%). Those who received training on SP and use of PPEs were less likely to develop nosocomial infections. Standard precautions training and regular provision of PPEs are recommended.