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Abstract Background The majority of gallbladder cancers (GBCs) are discovered incidentally during cholecystectomy for gallstone disease or upon routine postoperative pathological examination. Identifying simple and effective differential diagnostic markers for GBC is of great importance for primary care physicians and health screening practitioners. Methods This study included 151 patients diagnosed with GBC or benign gallbladder disease (BGD). Preoperative routine blood tests, biochemical parameters, and D-dimer levels were retrospectively reviewed. Relevant laboratory indices were analyzed and compared. Receiver operating characteristic (ROC) curve analysis was used to evaluate the discriminative power. Results Levels of D-dimer and its derivative indicators, such as monocyte-to-lymphocyte ratio × D-dimer (MLRD), neutrophil-to-lymphocyte ratio × D-dimer (NLRD), D-dimer-to-platelet ratio, the systemic immune-inflammation index × D-dimer (SIID), were significantly elevated in the GBC patients compared with the BGD patients (all P < 0.0001). The combination of D-dimer and C-reactive protein-to-albumin ratio demonstrated strong discriminatory efficacy, achieving an AUC of 0.846, with 80.00% sensitivity and 79.21% specificity ( P < 0.0001). Furthermore, the model combining MLRD, NLR, and SIID yielded an AUC of 0.752 ( P = 0.0003) for discriminating the early-stage from advanced-stage GBC, with a specificity of 92.86%. Conclusion D-dimer and its derivative indicators may possess independent diagnostic value in distinguishing GBC from BGD. The combination of MLRD, NLR and SIID may aid in differentiating between early-stage and advanced-stage GBC. These indicators represent a potentially simple and valuable preoperative screening tool, particularly for early GBC detection. These preliminary findings warrant confirmation in prospective studies with larger sample sizes.