为了建立适用于不确定性甲状腺肿瘤的靶向下一代测序（NGS）的最佳和最小侵入性诊断方法。患有不确定性甲状腺肿瘤的患者在单一三级医疗中心进行前瞻性招募和分析。我们对手术标本进行细针穿刺吸引（FNA）和核心针穿刺活检（CNB），以确认每个采样过程的质量。将FNA的细胞学诊断、CNB的组织学诊断和最终手术的确认诊断进行比较，以证明这些方法在不确定性甲状腺肿瘤中的一致性。分别评估FNA和CNB的样本质量，以确定靶向NGS的最佳方法。最后，我们对一例病例进行超声引导下的CNB和FNA（US-CNB和US-FNA）以确认临床可行性作为术前最小侵入性诊断方法。共招募了6名女性患者（平均年龄为50.83±15.18岁）患有不确定性甲状腺肿瘤（平均大小为1.79±0.91cm）进行进一步分析。在前五例中，CNB可以获得病理诊断，CNB样本的质量优于FNA，即使是在10倍稀释之后仍然如此。可以通过NGS检测与甲状腺恶性肿瘤相关的基因突变。在接受US-CNB治疗的病例中，成功获得了病理和靶向NGS结果，这表明了甲状腺恶性肿瘤可能性，从而促进了后续治疗的立即决策。CNB可以作为不确定性甲状腺肿瘤的最小侵入性诊断方法，通过提供病理诊断和合格样本，便于进行突变基因的检测，从而促进了适当和及时的治疗。©2023年作者（在Springer Nature的Springer-Verlag GmbH Germany的专属许可下）。

To establish the optimal and minimally invasive diagnostic approach for targeted next-generation sequencing (NGS) in the indeterminate thyroid tumors.The patients with indeterminate thyroid tumors were prospectively recruited and analyzed in a single tertiary medical center. We performed FNA and core needle biopsy (CNB) at the surgical specimens to confirm the quality of each sampling procedure. Cytological diagnosis by FNA, histological diagnosis by CNB and confirmed diagnosis by final surgery were compared to demonstrate the agreement among these approaches for the indeterminate thyroid tumors. The quality of the samples from FNA and CNB was evaluated, respectively to determine the optimal approach for targeted NGS. Finally, we performed ultrasound-guided CNB and FNA (US-CNB and US-FNA) on one case to confirm the clinical feasibility of being a pre-operative minimally invasive diagnostic approach.A total of 6 female patients (average age: 50.83 ± 15.18 years) with indeterminate thyroid tumors (average size: 1.79 ± 0.91 cm) were recruited for further analyses. The pathological diagnoses could be obtained by CNB in the first five cases, and the sample quality of CNB for targeted NGS was better than that of FNA, even after 10X dilution. The gene mutations associated with thyroid malignancy could be detected by NGS. In the case treated with US-CNB, the pathological and targeted NGS results were successfully obtained, which suggested the possibility of thyroid malignancy to facilitate immediate decision of subsequent treatment.CNB could serve as a minimally invasive diagnostic approach in the indeterminate thyroid tumors by providing pathological diagnoses and qualified samples for detection of mutated genes, which facilitates appropriate and immediate management.© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.