将 microRNA 标记物与下一代测序组合相结合可能会增强对细胞学不确定的甲状腺结节的风险评估。 ThyGeNEXT-ThyraMIRv1 多平台测试版本 1 在对 RAS 突变的不确定甲状腺结节进行风险分层方面的效用有限。我们试图在临床实践中验证更新的 ThyraMIRv2 平台。ThyGeNEXT/ThyraMIRv2（一种 3 层 microRNA 分类器）使用先前研究的 Bethesda III/IV 结节多机构队列进行评估，阳性结果具有恶性肿瘤风险≥10% 。此外，还评估了 ThyraMIRv2 在 RAS 突变的不确定性甲状腺结节中的临床效用。在 366 个不确定性甲状腺结节中，ThyraMIRv2 平台产生了 30.3% 的阳性检出率。与 ThyraMIRv2 平台结节相比，ThyraMIRv2 平台结节的手术率更高（63.9% vs 36.1%，P < .0001），具有乳头状核特征诊断的癌症/非浸润性滤泡性甲状腺肿瘤（65.9% vs 25.0%，P < .0001）。与多平台测试版本1相比，ThyraMIRv2平台的诊断测试参数没有明显改善。在 68 个 RAS 突变结节中，ThyraMIRv2 分别将 36.8%、55.9% 和 7.4% 分类为阳性、中度和阴性。所有中度结节的恶性风险≥10%，并与阳性队列合并。 RAS 突变阳性患者的手术率（81.0% vs 60.0%，P = .272）或具有乳头状核特征的癌症/非浸润性滤泡性甲状腺肿瘤诊断（47.6% vs 40.0%，P > .999）没有显着差异/温和和消极的群体。对于 RAS 突变结节，与 ThyGeNEXT-ThyraMIRv1 多平台测试版本 1 相比，ThyraMIRv2 表现出更高的敏感性（93.8% vs 64.7，P = .003）和降低的特异性（4.5% vs 34.8%，P = .008），并且具有可比的阴性预测结果值（33.3% vs 40.0%，P = .731）和阳性预测值（58.8% vs 59.5%，P = .864）。与 ThyGeNEXT-ThyraMIRv1 多平台测试版本 1 相比，ThyraMIRv2 平台不能改善不确定的甲状腺结节恶性肿瘤分层。 ThyraMIRv2 改善了恶性 RAS 突变结节的检测，但增加了误报。未来的研究需要包含更多具有手术病理学结果的 RAS 突变队列，以更好地表征该分类器的性能参数。版权所有 © 2024 Elsevier Inc. 保留所有权利。

Integrating microRNA markers with next-generation sequencing panels may enhance risk assessment of cytologically indeterminate thyroid nodules. The ThyGeNEXT-ThyraMIRv1 multiplatform test version 1 demonstrated limited utility in risk-stratifying RAS-mutated indeterminate thyroid nodules. We sought to validate the updated ThyraMIRv2 platform in clinical practice.ThyGeNEXT/ThyraMIRv2, a 3-tiered microRNA classifier, were evaluated using a previously studied multi-institutional cohort of Bethesda III/IV nodules, with positive results having risk of malignancy ≥10%. In addition, ThyraMIRv2's clinical utility in RAS-mutated indeterminate thyroid nodules was assessed.In 366 indeterminate thyroid nodules, ThyraMIRv2 platform yielded a 30.3% positive-call rate. ThyraMIRv2 platform + nodules had greater operative rates (63.9% vs 36.1%, P < .0001) and cancer/noninvasive follicular thyroid neoplasm with papillary-like nuclear features diagnosis (65.9% vs 25.0%, P < .0001) than ThyraMIRv2 platform nodules. Compared with multiplatform test version 1, ThyraMIRv2 platform's diagnostic testing parameters did not improve significantly. Among 68 RAS-mutated nodules, ThyraMIRv2 classified 36.8%, 55.9%, and 7.4% as positive, moderate, and negative, respectively. All moderate nodules had risk of malignancy ≥10% and were combined with the positive cohort. No significant differences existed in operative rate (81.0% vs 60.0%, P = .272) or cancer/noninvasive follicular thyroid neoplasm with papillary-like nuclear features diagnosis (47.6% vs 40.0%, P > .999) between RAS-mutated positive/moderate and negative groups. For RAS-mutated nodules, ThyraMIRv2 demonstrated improved sensitivity (93.8% vs 64.7, P = .003) and decreased specificity (4.5% vs 34.8%, P = .008) compared with ThyGeNEXT-ThyraMIRv1 multiplatform test version 1, with comparable negative predictive value (33.3% vs 40.0%, P = .731) and positive predictive value (58.8% vs 59.5%, P = .864).ThyraMIRv2 platform does not improve indeterminate thyroid nodule malignancy stratification compared to ThyGeNEXT-ThyraMIRv1 multiplatform test version 1. ThyraMIRv2 improves malignant RAS-mutated nodule detection but increases false positives. Future studies encompassing a larger cohort of RAS-mutated with surgical pathology results are warranted to better characterize the performance parameters of this classifier.Copyright © 2024 Elsevier Inc. All rights reserved.