Background: Current diagnostic methods, including tumor markers such as alpha-fetoprotein (AFP), carbohydrate antigen 19-9 (CA19-9), and carcinoembryonic antigen (CEA), have limitations in sensitivity and specificity, particularly in differentiating Hepatocellular carcinoma (HCC) from chronic liver diseases. This study investigated the diagnostic efficacy of these markers, individually and in combination, using chemiluminescence assay (CLIA) in patients with and without HCC. Methods: A cross-sectional study design was employed, analyzing data from 800 patients at Ibn Sina Diagnostic and Imaging Center in Dhaka. Tumor marker levels were assessed using CLIA kits, and associations with HCC diagnosis, tumor differentiation, occupation, age, and tumor size were examined. Results: Descriptive statistics revealed higher tumor marker levels in poorly differentiated tumors compared to welldifferentiated ones and controls. The combination of AFP, CA19-9, and CEA showed superior diagnostic accuracy for HCC, with a sensitivity of 86.5% and specificity of 92.3%. Occupation and age were found to correlate with tumor marker levels and HCC risk, with certain occupations and older age associated with larger tumor sizes. Regression analysis confirmed the associations between tumor markers and HCC. Conclusions: The study demonstrated the potential of AFP, CA19-9, and CEA as a panel of tumor markers for HCC detection and monitoring. CLIA emerged as a reliable diagnostic tool, offering high specificity and accuracy. Occupational factors and age were identified as relevant considerations in HCC risk assessment.