title: Automated quantitative RNA in situ hybridization for resolution of equivocal and heterogeneous ERBB2 (HER2) status in invasive breast carcinoma.
abstract: 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.
authors.affiliation: Department of Molecular Pathology, Cleveland Clinic, Cleveland, Ohio 44195, USA.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Wang
authors.forename: Bryce P
authors.initials: BP
authors.lastname: Portier
authors.forename: Aaron M
authors.initials: AM
authors.lastname: Gruver
authors.forename: Son
authors.initials: S
authors.lastname: Bui
authors.forename: Hongwei
authors.initials: H
authors.lastname: Wang
authors.forename: Nan
authors.initials: N
authors.lastname: Su
authors.forename: Hong-Thuy
authors.initials: HT
authors.lastname: Vo
authors.forename: Xiao-Jun
authors.initials: XJ
authors.lastname: Ma
authors.forename: Yuling
authors.initials: Y
authors.lastname: Luo
authors.forename: G Thomas
authors.initials: GT
authors.lastname: Budd
authors.forename: Raymond R
authors.initials: RR
authors.lastname: Tubbs
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001943:Breast Neoplasms / Q000175:diagnosis / Q000235:genetics* / Q000473:pathology*; D018567:Breast Neoplasms, Male / Q000175:diagnosis / Q000235:genetics / Q000473:pathology; D002277:Carcinoma / Q000175:diagnosis / Q000235:genetics* / Q000473:pathology*; D005260:Female; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D008875:Middle Aged; D060787:Neoplasm Grading; D009361:Neoplasm Invasiveness; D009367:Neoplasm Staging; D012333:RNA, Messenger; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics*; D012680:Sensitivity and Specificity
chemical_list: D012333:RNA, Messenger; D018719:Erb-b2 Receptor Tyrosine Kinases