Background: Somatic mutations of telomerase reverse-transcriptase (TERT) promoter and circulating microRNAs (miRNA) show potential biomarkers for several neoplastic diseases, including liver cancer. In this study, we evaluated the combinatory uses of TERT promoter mutations along with miRNA-122 expression level for screening HBV-related HCC. Methods: Nested PCR assay was optimised for detecting TERT promoter mutations C228T (-124G≥A) and C250T (-146G≥A) directly from patients’ blood. miR122 expression was quantified by real-time PCR. Diagnostic performance of combination between TERT promoter mutations and miR122 were assessed in 249 patients with HBV related liver diseases. Results: TERT promoter mutations could be identified at the detection limit of 1% by optimized PCR assay that helps detect C228T (-124G≥A) mutation in 22 HCC serum samples. Triple combined use of TERT promoter mutations, mir-122 and AFP levels (AFP@miR122Tert) obtained much better diagnostic performance to distinguish HCC from CHB (AUC = 0.97), LC (AUC = 0.93) and the group without HCC (LC + CHB, AUC = 0.96) compared to single uses. Notably, among HBV patients with AFP levels ≤ 20 ng/μl, the @miR122Tert panel also provided a better outcomes in discriminating HCC from the other conditions [CHB, AUC = 0.96; LC, AUC = 0.89, CHB and LC (CHB + LC), AUC = 0.94]. Conclusions: The triple combinatory use of circulating TERT promoter mutation, mir-122 and AFP is a potential biomarker panel that may be applied to improve diagnostics of HCC in HBV patients, especially in HBV-related LC patients with normal AFP levels.