Preoperative staging of suspicious axillary lymph nodes (ALNs) allows patients to be triaged to ALN dissection or to sentinel lymph node biopsy (SLNB). Ultrasound-guided fine needle aspiration (FNA) and cytology of ALN is moderately sensitive but its clinical utility relies heavily on the cytologist’s experience. We proposed that the 5-h automated GeneXpert system-based prototype breast cancer detection assay (BCDA) that quantitatively measures DNA methylation in ten tumor-specific gene markers could provide a facile, accurate test for detecting cancer in FNA of enlarged lymph nodes. We validated the assay in ALN-FNA samples from a prospective study of patients (N = 230) undergoing SLNB. In a blinded analysis of 218 evaluable LN-FNAs from 108 malignant and 110 benign LNs by histology, BCDA displayed a sensitivity of 90.7% and specificity of 99.1%, achieving an area under the ROC curve, AUC of 0.958 (95% CI: 0.928–0.989; P < 0.0001). Next, we conducted a study of archival FNAs of ipsilateral palpable LNs (malignant, N = 72, benign, N = 53 by cytology) collected in the outpatient setting prior to neoadjuvant chemotherapy (NAC). Using the ROC-threshold determined in the prospective study, compared to cytology, BCDA achieved a sensitivity of 94.4% and a specificity of 92.5% with a ROC-AUC = 0.977 (95% CI: 0.953–1.000; P < 0.0001). Our study shows that the automated assay detects cancer in suspicious lymph nodes with a high level of accuracy within 5 h. This cancer detection assay, scalable for analysis to scores of LN FNAs, could assist in determining eligibility of patients to different treatment regimens.

可疑腋窝淋巴结 (ALN) 的术前分期允许将患者分类为 ALN 清扫或前哨淋巴结活检 (SLNB)。超声引导下的细针穿刺 (FNA) 和 ALN 细胞学检查具有中等敏感性，但其临床实用性在很大程度上依赖于细胞学家的经验。我们提出，基于 GeneXpert 系统的 5 小时自动化原型乳腺癌检测试验 (BCDA) 可以定量测量十种肿瘤特异性基因标记中的 DNA 甲基化，可以为检测肿大淋巴结 FNA 中的癌症提供一种简便、准确的测试。我们在来自患者前瞻性研究的 ALN-FNA 样本中验证了该检测（N = 230) 正在接受 SLNB。在对来自 108 个恶性和 110 个良性 LN 的 218 个可评估 LN-FNA 的组织学盲法分析中，BCDA 显示出 90.7% 的敏感性和 99.1% 的特异性，实现了 ROC 曲线下面积，AUC 为 0.958（95% CI：0.928） –0.989；P  < 0.0001)。接下来，我们对 在新辅助化疗 (NAC) 之前在门诊环境中收集的同侧可触及 LN（恶性，N  = 72，良性，细胞学N = 53）的存档 FNA 进行了研究。使用前瞻性研究中确定的 ROC 阈值，与细胞学相比，BCDA 的敏感性为 94.4%，特异性为 92.5%，ROC-AUC = 0.977（95% CI：0.953–1.000；P < 0.0001)。我们的研究表明，自动化检测可在 5 小时内以高准确度检测可疑淋巴结中的癌症。这种癌症检测分析可扩展到分析 LN FNA 的分数，可以帮助确定患者是否适合不同的治疗方案。