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Droplet digital polymerase chain reaction detection of HER2 amplification in formalin fixed paraffin embedded breast and gastric carcinoma samples.

Authors
 Yazhen Zhu  ;  Dan Lu  ;  Maruja E. Lira  ;  Qing Xu  ;  Yunzhi Du  ;  Jianghong Xiong  ;  Mao Mao  ;  Hyun Cheol Chung  ;  Guangjuan Zheng 
Citation
 Experimental and Molecular Pathology, Vol.100(2) : 287-293, 2016 
Journal Title
 Experimental and Molecular Pathology 
ISSN
 0014-4800 
Issue Date
2016
MeSH
Adult ; Aged ; Aged, 80 and over ; Breast Neoplasms/diagnosis ; Breast Neoplasms/genetics* ; Breast Neoplasms/metabolism ; Cell Line, Tumor ; Female ; Fixatives/chemistry ; Formaldehyde/chemistry ; Gene Amplification ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Middle Aged ; Paraffin ; Polymerase Chain Reaction/methods* ; Receptor, ErbB-2/genetics* ; Receptor, ErbB-2/metabolism ; Reproducibility of Results ; Sensitivity and Specificity ; Stomach Neoplasms/diagnosis ; Stomach Neoplasms/genetics* ; Stomach Neoplasms/metabolism ; Tissue Embedding/methods ; Tissue Fixation/methods
Keywords
Breast cancer ; Droplet digital PCR ; Gastric cancer ; HER2 amplification
Abstract
RATIONALE: Human epidermal growth factor receptor 2 (HER2) is a key driver of tumorigenesis, and over-expression as a result of HER2 gene amplification has been observed in a number of solid tumors. Recently HER2 has become an important biomarker for the monoclonal antibody treatment of HER2-positive metastatic breast and advanced gastric cancer. The HER2 targeting antibody trastuzumab treatment requires accurate measurement of HER2 levels for proper diagnosis. Droplet digital PCR (ddPCR) with highly direct, precise and absolute nucleic acid quantification could be used to detect HER2 amplification levels. OBJECTIVES: Our objective was to evaluate a robust, accurate and less subjective application of ddPCR for HER2 amplification levels and test the assay performance in clinical formalin-fixed paraffin-embedded (FFPE) breast and gastric carcinoma samples. METHODS: Genomic DNA from HER2 amplified cell line SK-BR-3 was used to set up the ddPCR assays. The copy number of HER2 was compared to the chromosome 17 centromere reference gene (CEP17), expressed as HER2:CEP17 ratio. Genomic DNAs of FFPE specimens from 145 Asian patients with breast and gastric carcinomas were assayed using both standard methods, immunohistochemistry (IHC) and/or fluorescence in situ hybridization (FISH), and ddPCR. RESULTS: Based on 145 clinical breast and gastric carcinoma cases, our study demonstrated a high concordance of ddPCR results to FISH and IHC. In breast cancer specimens, the ddPCR results had high concordance with FISH and IHC defined HER2 status with a sensitivity of 90.9% (30/33) and a specificity of 100% (77/77). In gastric cancer specimens that were concordant in both FISH and IHC, our assay was 95.5% concordant with FISH and IHC (21/22). CONCLUSIONS: ddPCR has the advantage of automation and also allows levels of HER2 amplification to be easily evaluated in large numbers of samples, and presents a potential option to define HER2 status.
Full Text
http://www.sciencedirect.com/science/article/pii/S0014480015002385
DOI
10.1016/j.yexmp.2015.11.027
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Internal Medicine (내과학교실) > 1. Journal Papers
Yonsei Authors
정현철(Chung, Hyun Cheol) ORCID logo https://orcid.org/0000-0002-0920-9471
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URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/147091
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