Nurses play a crucial role in preventing and controlling transmission of the infection through the application of standard precautions and maintenance of the health care environment. In hospitals, infected patients are a source of infection transmission to other patients, health care workers, and visitors. Healthcare-related infections have a considerable impact on the morbidity and mortality rates in the intra- and extra-hospital environment, resulting in an increase in the time spent and costs of hospitalization, and are thus recognized as a serious world public health problem. For example, Nosocomial infection is one of the leading causes of death. The prevention and control of infections are critical for a well-functioning health system. The aim of this study is to evaluate the knowledge and practice of 60 nurses working in medical and surgical units at tertiary care hospital in relation to infection control measures. A questionnaire and an observation checklist were the data collection tool. The result of the study revealed that the majority of the sample had good knowledge about infection control measures, but they showed lack of practice in hand washing and using gloving which are the most significant items to prevent transmission of infection. In conclusion, nurse managers need to supervise the staff nurses on the practicing infection prevention standards and techniques and monitor nursing adherence to policies of the hospital. The Administrators should promote feedback on practice, individual reinforcement and appropriate rewards for the good practice.