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OBJECTIVE Evaluation of diagnostic accuracy of microRNA-21 (miR-21) in saliva and tumor tissue for presurgical assessment of lymph node metastasis in patients with oral squamous cell carcinoma (OSCC). STUDY DESIGN Unstimulated whole saliva and tumor tissue was obtained from clinically suspected patients with OSCC. A total of 130 patients diagnosed with OSCC were included as study participants. The assessment of cervical lymph node metastasis was done before surgery using imaging scans and post surgically confirmed by histopathologic examination of excised lymph nodes. miR-21 expression was evaluated using real-time polymerase chain reaction. The data was statistically analyzed for correlation analysis, cutoff values, sensitivity, and specificity. The κ statistic was applied to assess the degree of agreement between the lymph node metastasis and miR-21 expression. RESULTS miR-21 expression showed a statistically significant correlation with cervical lymph node metastasis with a diagnostic accuracy of 65% to 71.54% in saliva and 69% to 81.54% in tumor tissue. Very good agreement was observed between tumor tissue miR-21-3p and cervical lymph node metastasis with a specificity of 80.60% and a sensitivity of 82.40%. CONCLUSIONS miR-21 expression in saliva and tumor tissue of patients with OSCC showed high diagnostic accuracy for assessment of cervical lymph node metastasis. It can be used as an alternative for assessment of cervical lymph node metastasis before surgery.