This study was conducted on 25 healthy controls, 25 cirrhotic patients and 30 patients with HCC. All patients had been diagnosed with HCV-associated chronic liver disease. Methods: Serum PIVKAII and AFP were measured by enzyme linked immunosorbent assay (ELISA), while telomerase activity in peripheral blood was estimated by polymerase chain reaction- enzyme- linked immunosorbent assay (PCR- ELISA method). Results: Mean telomerase activity, PIVKAII and AFP levels were significantly higher in HCC patients as compared to both cirrhotic patients and controls, also a significant elevation in cirrhotic patients were found as compared to controls. Positive correlation was found between telomerase activity and size of hepatic focal lesions. Also, a positive correlation was found between both telomerase activity and PIVIKA-II and between the pathological grades of HCC. In HCC the sensitivity/specificity (88.2/79.6) of telomerase activity was much higher than both PIVIKA-II (80.5/69.3) and AFP (72.6/61.5). Conclusion: The usefulness of telomerase activity assay in HCC diagnosis and it's superiority to other tumor marker were recorded. Therefore, telomerase activity is a novel, available detector and prognostic marker for HCC diagnosis.