Introduction: Hemodialysis is a trusted intermediate procedure for management of chronic kidney disease (CKD) patients. As such CKD is an immune-deficient state, hence, they may be infected by hepatitis B virus (HCV) and hepatitis C virus (HCV) during hemodialysis. Eventually, HBV and HCV infection is an important cause of morbidity and mortality among patients of hemodialysis. Prevalence of HBV and HCV infection among hemodialysis patients is highly variable. Aim of the study: This study was designed to see the prevalence of HBV and HCV among hemodialysis patients of Tripura, India and to evaluate the mode of transmission. Materials and methods: This is a single center prospective study among the patients who has come for hemodialysis at the Dialysis Unit of Agartala Government Medical College and Govinda Ballav Pant Hospital during the period of January 2011 to June 2011. Results: Male predominance has been observed among the hemodialysis patients and mean age of the patients was 48.16 years. New HBV infection was found in 5.5% of the hemodialysis patients (cumulative 7.3%) and 10.9% (cumulative 12.12%) of patients got new HCV infection during hemodialysis. Coinfection with both HBV and HCV was found among 1.2% of hemodialysis patients. History of blood transfusion was found in 100% of newly infected HBV and HCV patients. Discussion: The evaluation suggests that blood transfusion may be the single most important risk factor for HBV and HCV infection among hemodialysis patients. Newer methods of screening of blood in blood bank like nucleic acid test or polymerase chain reaction amplification of nucleic acids should be adopted to prevent HBV and HCV transmission