BACKGROUND An estimated 130-170 million people worldwide are chronically infected with HCV.(1) In Europe the highest prevalence of HCV infections is in the IDU population.(2) As traditional HCV screening relies on the detection of HCV antibody or HCV RNA in blood, screening in high-risk groups such as IDU is difficult due to poor venous access caused by damaged veins. OBJECTIVE In this study DBS was evaluated as an alternative sample type to blood for the detection of HCV RNA. METHODS The endpoint detection limit, inter-assay and intra-assay variability of the method were determined. The DBS method was compared to our routine frontline assay using a panel of paired DBS and blood samples. The effect of different storage temperatures and length of storage time on the stability of HCV RNA in DBS was also assessed. RESULTS The endpoint detection limit of the method based on results from mock DBS was 250 IU/ml. The method was shown to be precise and robust. The sensitivity and specificity of the method was found to be 100% and 95.8%, respectively. No significant variation in the stability of HCV RNA in DBS over a 1 year period at a range of different temperatures was observed. CONCLUSIONS A sensitive and stable method was developed for the detection of HCV RNA in DBS. Screening high-risk populations using DBS as a sample type may improve uptake of HCV testing by increasing opportunity for patients to be tested and consequently increasing access to treatment.