Objective: To explore the role of telomerase activity detected in biopsy samples for evaluating the efficacy of lapa- roscopic radiofrequency ablation (RFA) therapy in patients with hepatocellular carcinoma (HCC) and liver cirrhosis. Methods: From August 2001 to October 2004, 34 cirrhotic patients with HCC were treated by laparoscopic RFA under general anesthe- sia. A total of 34 tumors, with a mean maximum tumor diameter of 4.0 ± 1.0 cm, were all located on the liver surface or adja- cent to the gallbladder. Laparoscopic ultrasound-guided core biopsy for liver lesions was performed before and immediately after RFA therapy. In these biopsy samples, telomerase activity was detected by the ELISA-based telomeric repeat amplifica- tion protocol (ELISA-TRAP) assay, and pathological examination was routinely performed. Results: Laparoscopic RFA was successfully performed in all the 34 patients. A complete tumor necrosis was achieved in all patients on the contrast-enhanced helical CT scanning one month after laparoscopic RFA. The positive rates of telomerase activity and histopathologic diagnosis in biopsy samples were 91.2% (31/34) and 100% (34/34) respectively before RFA, and 26.5% (9/34) and 0% respectively after RFA. During a median follow-up period of 35 months (range, 18–51 months), the rates of local tumor recurrence at the ablation sites in post-RFA telomerase-positive and negative patients were 88.9% (8/9) and 4% (1/25) respectively (P < 0.01), and the rates of distant recurrence within the livers were 0% (0/9) and 12% (3/25) respectively (P > 0.05). Conclusion: For cirrhotic patients with HCC treated by laparoscopic RFA, detection of telomerase activity in biopsy samples may be useful for evaluating the therapeutic efficacy of RFA and predicting postoperative local tumor recurrence.