Background: The rising incidence of vertebral body fractures, vertebral infections and metastatic disease increases the need for diagnostic modalities with high specificity. Biopsy remains essential, yet comparative data on CT-guided versus intraoperative percutaneous fluoroscopy-guided biopsy are limited. Methods: This retrospective study compared two cohorts biopsied for spinal lesions between April 2015 and April 2024: CT-guided biopsy (n = 62), and intraoperative percutaneous biopsy (n = 154). Groups were analyzed for demographic and clinical characteristics, and diagnostic yield was defined by the conclusiveness of the primary biopsy; statistical comparisons were performed using Fisher’s exact test. Results: CT-guided biopsy yielded conclusive results in 46 of 62 cases (74.2%), whereas intraoperative, fluoroscopy-guided biopsy was conclusive in 41 of 154 cases (26.6%), representing a statistically significant difference (p < 0.001). In analogy, propensity score matching (1:1) resulted in similar significant (p < 0.001) results (CT-guided group vs. intraoperative, fluoroscopy-guided group: 86.7% vs. 35.6%) Conclusions: CT-guided biopsy demonstrated a substantially higher rate of conclusive results compared with intraoperative biopsy in this cohort. Further studies with larger and more balanced cohorts are needed to strengthen clinical recommendations.
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