Automated Quantitative RNA in Situ Hybridization for Resolution of Equivocal and Heterogeneous ERBB2 (HER2) Status in Invasive Breast Carcinoma
Automated Quantitative RNA in Situ Hybridization for Resolution of Equivocal and Heterogeneous ERBB2 (HER2) Status in Invasive Breast Carcinoma
复制标题
DOI：
10.1016/j.jmoldx.2012.10.003
复制
发表时间：
2013-03-01
影响因子：
4.1
通讯作者：
Tubbs, Raymond R.
中科院分区：
文献类型：
--
作者：
Wang, Zhen;Portier, Bryce P.;Tubbs, Raymond R.
基于乳腺癌中HER 2状态的患者管理是个性化医疗的典型示例，但仍受到不明确的检测和瘤内异质性的阻碍。我们开发了一种全自动、定量、明视野原位杂交技术（RNAscope），应用其定量132例浸润性乳腺癌中的单细胞HER 2 mRNA水平，并将结果与实时定量PCR（qPCR）和食品和药物管理局批准的方法（包括荧光原位杂交（FISH），IHC，显色原位杂交，和双重原位杂交。在FISH结果明确的病例中，RNAscope和qPCR与FISH的一致率均为97.3%。在肿瘤内异质性或FISH结果不明确的病例中，RNAscope上级qPCR。这种新的检测方法可以实现最终的HER 2状态分辨率，作为当前检测算法的反射检测。在单细胞水平的定量原位RNA测量可以广泛地应用于伴随诊断应用。（J Mol Diagn 2013，15：210-219; http：//dx.doi.org/10.1016/j.jmoldx.2012.10.003）
Patient management based on HER2 status in breast carcinoma is an archetypical example of personalized medicine but remains hampered by equivocal testing and intratumoral heterogeneity. We developed a fully automated, quantitative, bright-field in situ hybridization technique (RNAscope), applied it to quantify single-cell HER2 mRNA levels in 132 invasive breast carcinomas, and compared the results with those by real-time quantitative PCR (qPCR) and Food and Drug Administration-approved methods, including fluorescence in situ hybridization (FISH), IHC, chromogenic in situ hybridization, and dual in situ hybridization. Both RNAscope and qPCR were 97.3% concordant with FISH in cases in which FISH results were unequivocal. RNAscope was superior to qPCR in cases with intratumoral heterogeneity or equivocal FISH results. This novel assay may enable ultimate HER2 status resolution as a reflex test for current testing algorithms. Quantitative in situ RNA measurement at the single-cell level may be broadly applicable in companion diagnostic applications. (J Mol Diagn 2013, 15: 210-219; http://dx.doi.org/10.1016/j.jmoldx.2012.10.003)