Introduction: Health care associated infections (HAIs) are considered as major patient safety concerns that cause avoidable excess morbidity and mortality among hospitalized patients. It is prolonged hospital stay, increase resistant to life-saving medications, drive up the cost of medical services, create long term disabilities that reduce quality of life and may lead to death. As well as human cost, healthcare systems carry massive additional financial burden. Aim of study: This study conducted to determine promoting and impeding factors related to infection prevention and control program through system analysis, hand hygiene compliance between health care workers (HCWs) in addition to assess knowledge, attitude and practices of the medical staff regarding standard precautions of infection prevention and control in the regular ward of Khartoum Teaching Hospital (KTH). Methods: A cross section study in which system analysis was done through in-depth personal interviews with four key personnel and also by observation using checklists to assess structure of the twenty regular wards of the hospital and hand hygiene compliance between HCWs. Knowledge, attitude and practices were assessed using self-administrated questionnaire which distributed to 136 doctors and 145 nurses. Results: Absence of structured surveillance system makes it difficult to estimate the magnitude of the problem, but it is vary from common and very common according to most provider consideration. General compliance with hand hygiene is 30.5% between HCWs and these much less in surgical wards (21.6%) than medical wards (43.4%) (P≤0.001). Only 20.2% of medical staff received training regarding IPC and this much better between nurses (31.7%) than doctors (8.8%) (P≤0.001). Conclusion: Presence of full-time working ICT, available national guidelines for IPC and recognition of most HCWs with their general role in prevention of HAIs are the main promoting factors. While Absence of supervision and support for infection control team (ICT), absence of specific budget that directed to IPC program and ineffective Surveillance system are the main impeding factors. Recommendation: 1) Raise leadership commitment and support. 2) Allocate specific running budget for IPC program. 3) Reset-up of surveillance system. 4) Organize regular training courses.