e16622 Background: Many cancers could be driven by some specially mutations exampled as EGFR in lung cancer, influenced prognosis prominently. However, there were few related studies in liver cancer, although primary liver cancer was a common malignant tumor causing high morbidity and mortality. Methods: 97 patients diagnosed with primary hepatocellular carcinoma were enrolled in this study. Operative tissue samples were collected and analyzed using hybridization capture based NGS ERSeq method from all patients. Results: 1088 somatic mutations were identified in tissue samples. Targeted-capture sequencing of 1,021 genes that are frequently mutated in solid tumors revealed that the most frequently mutated genes were TP53 (59%), TERT (32%) and AXIN1 (19%). By using the least absolute shrinkage and selection operator (LASSO), the recurrence risks were constructed on the basis of five-gene mutations, AXIN1, CTNNB1, LRP1B, PDGFRA and TP53. The ROC curve was 0.813 and 0.882 in training and validation cohorts, respectively. TP53 R249G/S point mutation was predicted prognostic independently. Up to 60% TP53 R249S mutated patients occurred recurrence, while less than 30% TP53 R249 wildtype patients occurred recurrence ( P = 0.0002). Patients with R249G/S point mutation showed a worse prognosis. The median disease-free survival time was 8.0 VS 45.0 months in R249G/S mutated group (n = 79) and R249- wildtype (n = 18) group, respectively (Log-rank test, P = 0.0364). Conclusions: In various cancer types, TP53 has a broad-spectrum mutation, but only in hepatocellular carcinoma, TP53 mutations are highly concentrated at the R249 point, which was considered to be related to aflatoxin infection, and mutations at this site can significantly affect the prognosis of patients. The discovery of this target also provide clue for subsequent targeted treatment of liver cancer.