title: Quantitative real-time polymerase chain reaction is an alternative method for the detection of HER-2 amplification in formalin-fixed paraffin-embedded breast cancer samples.
abstract: Fluorescent in situ hybridization (FISH) and immunohistochemistry (IHC) are the most common methods that are used to quantify HER-2 gene and protein levels, respectively, in human breast cancer. However, due to bad sample quality, some samples are unable to be subjected to a FISH assay. We evaluated 71 formalin-fixed paraffin-embedded (FFPE) breast carcinoma specimens by quantitative real-time polymerase chain reaction (qPCR), IHC, and FISH. We also performed qPCR and FISH assays on delayed formalin-fixed (DDF) samples. The qPCR results were in complete concordance with the results of IHC and FISH. In regards to the DDF samples, the HER-2 fluorescent signal seemed decayed compared with that of the DDF samples after 1 h. However, the qPCR method still works well up to 12 hours. Our results indicated that qPCR was obviously superior to FISH in cases that were not fixed in a reasonable amount of time. However, qPCR can be an alternative method by which to perform HER2 amplification assays in breast cancer.
authors.affiliation: Laboratory of Pathology, West China Hospital, Sichuan University Sichuan, China ; Department of Pathology, West China Hospital, Sichuan University Sichuan, China.
authors.forename: Tianjie
authors.initials: T
authors.lastname: Pu
authors.affiliation: Laboratory of Pathology, West China Hospital, Sichuan University Sichuan, China.
authors.forename: Peng
authors.initials: P
authors.lastname: Guo
authors.affiliation: Laboratory of Pathology, West China Hospital, Sichuan University Sichuan, China ; Department of Pathology, West China Hospital, Sichuan University Sichuan, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Qiu
authors.affiliation: Laboratory of Pathology, West China Hospital, Sichuan University Sichuan, China.
authors.forename: Shinan
authors.initials: S
authors.lastname: Chen
authors.affiliation: Laboratory of Pathology, West China Hospital, Sichuan University Sichuan, China ; Department of Pathology, West China Hospital, Sichuan University Sichuan, China.
authors.forename: Libo
authors.initials: L
authors.lastname: Yang
authors.affiliation: Department of Pathology, West China Hospital, Sichuan University Sichuan, China.
authors.forename: Linyong
authors.initials: L
authors.lastname: Sun
authors.affiliation: Laboratory of Pathology, West China Hospital, Sichuan University Sichuan, China.
authors.forename: Feng
authors.initials: F
authors.lastname: Ye
authors.affiliation: Laboratory of Pathology, West China Hospital, Sichuan University Sichuan, China ; Department of Pathology, West China Hospital, Sichuan University Sichuan, China.
authors.forename: Hong
authors.initials: H
authors.lastname: Bu
mesh_terms: D014408:Biomarkers, Tumor / Q000032:analysis / Q000235:genetics*; D001943:Breast Neoplasms / Q000201:enzymology / Q000235:genetics* / Q000473:pathology; D005260:Female; D005404:Fixatives; D005557:Formaldehyde; D005784:Gene Amplification; D015971:Gene Expression Regulation, Enzymologic; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007150:Immunohistochemistry; D017404:In Situ Hybridization, Fluorescence; D061986:MCF-7 Cells; D016612:Paraffin Embedding; D011237:Predictive Value of Tests; D060888:Real-Time Polymerase Chain Reaction; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000032:analysis / Q000235:genetics*; D015203:Reproducibility of Results; D013997:Time Factors; D016707:Tissue Fixation / Q000379:methods*; D057188:Workflow
chemical_list: D014408:Biomarkers, Tumor; D005404:Fixatives; D005557:Formaldehyde; D018719:Erb-b2 Receptor Tyrosine Kinases; C508053:ERBB2 protein, human
keywords: FISH; HER-2 copy number variation; IHC; breast neoplasm; delayed formalin-fixed; quantitative real-time polymerase chain reaction