Introduction: HCV infection in hemodialysis units is a significant cause of morbidity and mortality. The risk of HCV infection among dialysis patients is higher compared to the general population due to high potential blood exposures in hemodialysis settings. Multiple transfusions and hemodialysis are associated with complications, such as hepatitis B and C and certain sexually transmitted disease like, HIV infections. This study aimed to determine the prevalence of hepatitis C in multiple hemodialysis patients. Materials and methods: This study was carried at the Department of Microbiology, Shri. M. P. Shah Govt. Medical College and Guru Govind Govt. hospital, Jamnagar from 1 st June 2016 to 31st May 2017. Patients with kidney failure or end stage renal disease who received multiple hemodialysis were included in this study. Result: Serum samples from 229 patients are collected which including 134(58.5%) male and 95 (41.5%) female, were tested using enzyme-linked immune sorbent assay kits for hepatitis C infection. Out of them 43 (18.8%) were reactive for hepatitis C virus antibodies. Conclusion: Sensitive screening tests for hepatitis C infection and strict guidelines for patients with renal failure or end stage renal disease who received multiple hemodialysis are needed for prevention of transfusion related.