HER2DX, a multianalyte genomic test, has been clinically validated to predict breast cancer recurrence risk (relapse risk score), the probability of achieving pathological complete response post-neoadjuvant therapy (pCR likelihood score), and individual messenger RNA (mRNA) expression levels in patients with early-stage human epidermal growth factor receptor 2 (HER2)-positive breast cancer. This study delves into the comprehensive analysis of HER2DX’s analytical performance. Precision and reproducibility of HER2DX risk, pCR, and mRNA scores were assessed within and between laboratories using formalin-fixed paraffin-embedded (FFPE) tumor tissues and purified RNA. Robustness was appraised by analyzing the impact of tumor cell content and protocol variations including different instruments, reagent lots, and different RNA extraction kits. Variability was evaluated across intratumor biopsies and genomic platforms [RNA sequencing (RNAseq) versus nCounter], and according to protocol variations. Precision analysis of 10 FFPE tumor samples yielded a maximal standard error of 0.94 across HER2DX scores (1-99 scale). High reproducibility of HER2DX scores across 29 FFPE tumors and 20 RNAs between laboratories was evident (correlation coefficients >0.98). The probability of identifying score differences >5 units was ≤5.2%. No significant variability emerged based on platform instruments, reagent lots, RNA extraction kits, or TagSet thaw/freeze cycles. Moreover, HER2DX displayed robustness at low tumor cell content (10%). Intratumor variability across 212 biopsies (106 tumors) was <4.0%. Concordance between HER2DX scores from 30 RNAs on RNAseq and nCounter platforms exceeded 90.0% (Cohen’s κ coefficients >0.80). The HER2DX assay is highly reproducible and robust for the quantification of recurrence risk, pCR likelihood, and mRNA expression in early-stage HER2-positive breast cancer.

中文翻译：

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HER2DX 基因组测试对早期 HER2 阳性乳腺癌的分析验证

HER2DX 是一种多分析物基因组测试，经过临床验证，可以预测乳腺癌复发风险（复发风险评分）、新辅助治疗后实现病理完全缓解的概率（pCR 可能性评分）以及个体信使 RNA (mRNA) 表达水平。早期人类表皮生长因子受体 2 (HER2) 阳性乳腺癌患者。本研究深入探讨了 HER2DX 分析性能的综合分析。使用福尔马林固定石蜡包埋 (FFPE) 肿瘤组织和纯化的 RNA 在实验室内部和实验室之间评估 HER2DX 风险、pCR 和 mRNA 评分的精度和重现性。通过分析肿瘤细胞含量和方案变化（包括不同仪器、试剂批次和不同 RNA 提取试剂盒）的影响来评估稳健性。根据协议变化，跨肿瘤内活检和基因组平台[RNA测序（RNAseq）与nCounter]评估变异性。对 10 个 FFPE 肿瘤样本的精密分析得出 HER2DX 评分（1-99 等级）的最大标准误差为 0.94。实验室之间 29 个 FFPE 肿瘤和 20 个 RNA 的 HER2DX 评分具有明显的高再现性（相关系数 >0.98）。识别分数差异 >5 个单位的概率≤5.2%。根据平台仪器、试剂批次、RNA 提取试剂盒或 TagSet 解冻/冷冻循环，没有出现显着的差异。此外，HER2DX 在低肿瘤细胞含量 (10%) 下表现出稳健性。 212 个活检组织（106 个肿瘤）的肿瘤内变异性 <4.0%。 RNAseq 和 nCounter 平台上 30 个 RNA 的 HER2DX 分数之间的一致性超过 90.0%（Cohen's κ 系数 >0.80）。 HER2DX 检测具有高度重复性和稳健性，可用于量化早期 HER2 阳性乳腺癌的复发风险、pCR 可能性和 mRNA 表达。