Objective: To discuss the differential diagnostic value of fine needle aspiration biopsy (FNAB) combined with BRAF-V600E gene detection on benign and malignant thyroid nodules. Methods: 101 thyroid nodules obtained by routine pathological results of surgical operation after conducting retrospective analysis were diagnosed by preoperative ultrasound diagnosis as suspicious malignant nodules. At the same time, fine needle aspiration biopsy (FNAB) and BRAF-V600E gene detection were performed. With postoperative routine pathological results as the gold standard, the diagnostic value of using two different methods separately and jointly for suspicious malignant thyroid nodules were evaluated. Results: Among 101 suspicious malignant thyroid nodules of 100 patients' with postoperative pathological results, 21were benign nodules and 80 were malignant nodules. There were 75 BRAF-V600E mutant nodules, of which 70 were positive and 5 were negative by FNAB. There were 26 wild-type nodules of the BRAF-V600E gene, of which 9 were positive and 17 were negative by FNAB.The sensitivity and specificity of FNAB for suspicious malignant thyroid nodules were: 93.7%, 80.95%. The sensitivity and specificity of BRAF-V600E gene detection for suspicious malignant thyroid nodules were: 92.50 %, 95.24%. The sensitivity and specificity of FNAB combined with BRAF-V600E gene detection for suspicious malignant thyroid nodules were: 98.59%、100%. Conclusion: FNAB combined with BRAF-V600E gene detection can improve the efficiency of differential diagnosis of suspicious benign and malignant thyroid nodules. It has particularly higher specificity for papillary thyroid carcinoma, thus deserving clinical popularization and application. Keywords: Fine needle aspiration biopsy; Thyroid nodule; BRAF-V600Egene detection