Background: Occupational exposure from hepatitis C virus care is a substantial source of infection by blood borne pathogens among health care workers. Aim of the study: Assess nurses' knowledge and attitudes regarding hepatitis C virus. Study design: Multi-center study was carried out as a cross-sectional survey based on a self-administered questionnaire. It was conducted on (700) nurses who attended the central-hospital, Health Affairs, Al-humiate, Reproductive Health Hospital and Nursing School at Manfalout city and both male and females. Tools: Data were collected by using; self-administer questionnaire sheet: It contained demographic data and assessment of nurses' knowledge about HCV and assessment of nurses' attitude regarding HCV (Likert scale). Results: The majority of nurses (92.9%) had good level of knowledge about hepatitis C virus. The majority of the study nurses (96.3%) had positive attitude toward patient with hepatitis C virus. There was no a statistical significance difference between demographic data and nurses' attitude regarding patients with hepatitis C virus p. value (1.033*). On other hand, there was a statistical significance difference between demographic data and nurses' knowledge regarding patients with hepatitis C virus. Conclusion: There were positive and significant correlation among total score of nurses' knowledge and their attitude toward patients with hepatitis C virus (r. 0.146 and p. value 0.001). Recommendations: Continuing education programs on infection control measures are needed for nurses to increase their level of knowledge regarding patients with hepatitis C virus.