Hemodialysis (HD) patients are at increased risk for both hepatitis C virus (HCV) and hepatitis B virus (HBV) infections associated with contaminated blood and blood product transfusion and exposure to contaminated HD equipment during treatment. Thus, the present study aimed to assess the prevalence of HBV, HCV and HBV/HCV co-infection among HD-patients in the most common main three dialysis units in Ismailia governorate, Egypt. Furthermore, to find out the impact of HCV on the HBV infection in those HD-patient. This was done using both serological ELISA and real time PCR molecular techniques. The results showed that HCV infection was the most prevalent one representing about 26%, the HBV infection was less prevalent than HCV 9.3%, and the dual infection was rare representing only 2% of the studied HD-patients in Ismailia governorate. In addition, there was a non-significant difference of both HBV incidence and viral load (copies/mL) between the studied HD-patients with and without HCV infection [P-value=0.36].