Breast Cancer Subtype Classification Using 4-Plex Droplet Digital PCR
Breast Cancer Subtype Classification Using 4-Plex Droplet Digital PCR
复制标题
使用 4 重微滴数字 PCR 进行乳腺癌亚型分类
DOI：
10.1373/clinchem.2019.302315
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发表时间：
2019-08-01
影响因子：
9.3
通讯作者：
Wang, Jidong
中科院分区：
文献类型：
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作者：
Chen, Wenwen;Zheng, Jiaying;Wang, Jidong
背景：浸润性导管癌（IDCA）是最常见的浸润性乳腺癌。免疫组织化学 (IHC) 广泛用于分析雌激素受体 1 (ER)、孕激素受体 (PR) 和人表皮生长因子受体 2 (HER2)，有助于对肿瘤进行分类以指导治疗。 IHC 检查需要经验丰富的病理学家提供主观解释，从而降低了基于 IHC 的诊断的可重复性。在本研究中，我们开发了一种 4 重微滴数字 PCR (ddPCR)，用于同时定量分析福尔马林固定石蜡包埋 (FFPE) 样品中雌激素受体 1 (ESR1)、孕激素受体 (PGR)、erb-b2 受体酪氨酸激酶 2 (ERBB2) 和 pumilio RNA 结合家族成员 1 (PUM1) 的表达水平。 4 重 ddPCR 的重现性和线性动态范围。我们应用这种方法分析了来自乳腺 IDCA 患者的 95 份 FFPE 样本，并评估了 ddPCR 和 IHC 之间的一致性，以评价其对乳腺癌亚型进行分类的潜力。 结果：ERBB2、ESR1、PGR 和 PUM1 的每次反应的定量限 (LOQ) 分别为 25、50、50 和 50 个拷贝。 ESR1、PGR 和 PUM1 的动态范围从每个反应 50-1600 个拷贝扩展至每个反应 50-1600 个拷贝，ERBB2 的动态范围从每个反应 25 个拷贝扩展至 1600 个拷贝。 ERBB2、ESR1 和 PGR 的 4 重 ddPCR 和 IHC 之间的一致性相关系数分别为 96.8%、91.5% 和 85.1%。 ERBB2、ESR1 和 PGR 的曲线值下的受试者工作特征曲线面积分别为 0.991、0.977 和 0.920。结论：本研究中通过 4 重 ddPCR 对乳腺癌生物标志物状态的评估与 IHC 高度一致。 (C) 2019年美国临床化学协会
BACKGROUND: Infiltrating ductal carcinoma (IDCA) is the most common form of invasive breast cancer. Immunohistochemistry (IHC) is widely used to analyze estrogen receptor 1 (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2) that can help classify the tumor to guide the medical treatment. IHC examinations require experienced pathologists to provide interpretations that are subjective, thereby lowering the reproducibility of IHC-based diagnosis. In this study, we developed a 4-plex droplet digital PCR (ddPCR) for the simultaneous and quantitative analyses of estrogen receptor 1 (ESR1), progesterone receptor (PGR), erb-b2 receptor tyrosine kinase 2 (ERBB2), and pumilio RNA binding family member 1 (PUM1) expression levels in formalin-fixed paraffin-embedded (FFPE) samples.METHODS: We evaluated the sensitivity, reproducibility, and linear dynamic range of 4-plex ddPCR. We applied this method to analyze 95 FFPE samples from patients with breast IDCA and assessed the agreement rates between ddPCR and IHC to evaluate its potential in classifying breast cancer subtypes.RESULTS: The limits of quantification (LOQ) were 25, 50, 50, and 50 copies per reaction for ERBB2, ESR1, PGR, and PUM1, respectively. The dynamic ranges of ESR1, PGR, and PUM1 extended over 50-1600 copies per reaction and those of ERBB2 from 25 to 1600 copies per reaction. The concordance correlation coefficients between 4-plex ddPCR and IHC were 96.8%, 91.5%, and 85.1% for ERBB2, ESR1, and PGR, respectively. Receiver operating characteristic curve area under the curve values of 0.991, 0.977, and 0.920 were generated for ERBB2, ESR1, and PGR, respectively.CONCLUSIONS: Evaluation of breast cancer biomarker status by 4-plex ddPCR was highly concordant with IHC in this study. (C) 2019 American Association for Clinical Chemistry