It is known that Egypt has the largest burden of hepatitis C virus (HCV) infection in the world; and the prevalence of the virus reached 48% and as high risk as 87.5% among hemodialysis (HD) patients. Compliance with HD precautions implemented by HD nursing staff is essential in preventing the transmission of the virus in hemodialysis units (HDUs). The study aimed to assess nursing staffs knowledge and practice regarding prevention of HCV transmission in HDUs in a University Hospital in Egypt.