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Background and Aims: Telomerase activity and human telomerase reverse transcriptase (TERT), one of the major catalytic subunits of telomerase, have been reported as a marker of tumor aggressiveness and poor prognosis in some kinds of malignancies. However, no previous reports have shown a correlation between telomerase or TERT expression and patient outcome in ampullary carcinoma. In the present study, we examined telomerase activity and TERT expression in ampullary carcinoma to determine whether these parameters can be used as prognostic indicators of patient outcome.Methods: A total 46 ampullary carcinoma patients who underwent pancreatoduodenectomy with regional lymph node dissection were enrolled in this study. Telomerase activity was analyzed by the telomeric repeat amplification protocol assay using frozen tissue samples. TERT was examined by immunohistochemistry using paraffin embedded samples. Relationships between survival and clinicopathological factors including tumor characteristics, telomerase activity, and TERT expression were evaluated using univariate and multivariate analysis.Results: Telomerase activity was detected in 42 (91.3%) of ampullary carcinoma and 27 (58.7%) showed high activity (>8.9 TPG units, which is the maximum level of telomerase in normal duodenal tissue samples). In contrast, TERT was expressed in 35 (76.1%), including 21 weak and 14 strong immunohistochemical staining. Univariate analysis revealed that tumor depth (P<0.001), nodal status (P=0.013), UICC stage (P=0.009), perineural invasion (P<0.001), telomerase activity (P=0.031) were significantly associated with disease specific survival. TERT expression showed a tendency toward association with prognosis (P=0.054). In multivariate analysis, only telomerase activity remained independent predictors of prognosis (P=0.043). The 5-year survival rates of the patients with high or low telomerase activity were 66.2% and 94.4%, respectively.Conclusions: Our results suggest that telomerase activity has a prognostic significance for survival in patients with resected ampullary carcinoma. Telomerase activity may provide a new marker for evaluating the progression of patients with ampullary carcinoma, and may help to identify patients in need of adjuvant treatment.