Background: Hepatitis C virus (HCV) is a major public health problem globally. Diagnosis of HCV infection is mainly based on the detection of anti-HCV antibodies by the Enzyme Linked Immunosorbent Assay (ELISA) or Rapid Diagnostic Tests (RDTs) and HCV Ribonucleic Acid (RNA) by Polymerase Chain Reaction (PCR) of serum or plasma samples. Aims & objective: This study evaluate the efficacy of the RDT/third generation ELISA for detection of antibodies to hepatitis C virus (anti-HCV) in comparison with reverse transcriptase polymerase chain reaction (RT-PCR) to detect HCV RNA for the diagnosis of hepatitis C virus. Material & method: This was a comparative observational study. A total of 500 samples received were tested for HCV infection. Different techniques for diagnosis of HCV were used rapid card test (RDT), ELISA, and RT-PCR etc RDT/ELISA positive samples further tested for detection of HCV RNA by RT-PCR. HCV viral load was also measured. Results: Out of total positive cases majority of them (54.6%) were 21-30 years age group, predominantly male (77.3%).blood transfusion was the most common risk factor of HCV infection. Out of total 22 (4.4%) samples were reported positive for anti HCV antibody by RDT and ELISA, same number of samples positive for HCV RNA by RT-PCR. Conclusion: RT-PCR is very important gold standard test for early diagnosis of HCV infection as well as for monitoring antiviral therapy response.