PMID- 26626802
OWN - NLM
STAT- MEDLINE
DCOM- 20160906
LR  - 20160407
IS  - 1096-0945 (Electronic)
IS  - 0014-4800 (Linking)
VI  - 100
IP  - 2
DP  - 2016 Apr
TI  - Droplet digital polymerase chain reaction detection of HER2 amplification in 
      formalin fixed paraffin embedded breast and gastric carcinoma samples.
PG  - 287-93
LID - S0014-4800(15)00238-5 [pii]
LID - 10.1016/j.yexmp.2015.11.027 [doi]
AB  - 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.
CI  - Copyright (c) 2015 Elsevier Inc. All rights reserved.
FAU - Zhu, Yazhen
AU  - Zhu Y
AD  - Guangdong Provincial Hospital of Traditional Chinese Medicine (GPHTCM), 
      Guangzhou, China.
FAU - Lu, Dan
AU  - Lu D
AD  - Translational Bioscience and Diagnostics, WuXi AppTec Co., Ltd., Waigaoqiao Free 
      Trade Zone, Shanghai, China.
FAU - Lira, Maruja E
AU  - Lira ME
AD  - Pfizer Oncology, San Diego, California, United States.
FAU - Xu, Qing
AU  - Xu Q
AD  - Translational Bioscience and Diagnostics, WuXi AppTec Co., Ltd., Waigaoqiao Free 
      Trade Zone, Shanghai, China.
FAU - Du, Yunzhi
AU  - Du Y
AD  - Translational Bioscience and Diagnostics, WuXi AppTec Co., Ltd., Waigaoqiao Free 
      Trade Zone, Shanghai, China.
FAU - Xiong, Jianghong
AU  - Xiong J
AD  - Translational Bioscience and Diagnostics, WuXi AppTec Co., Ltd., Waigaoqiao Free 
      Trade Zone, Shanghai, China.
FAU - Mao, Mao
AU  - Mao M
AD  - Translational Bioscience and Diagnostics, WuXi AppTec Co., Ltd., Waigaoqiao Free 
      Trade Zone, Shanghai, China. Electronic address: mao_m@yahoo.com.
FAU - Chung, Hyun Cheol
AU  - Chung HC
AD  - Yonsei Cancer Center, Yonsei University College of Medicine, Seoul, South Korea. 
      Electronic address: UNCHUNG8@yuhs.ac.
FAU - Zheng, Guangjuan
AU  - Zheng G
AD  - Guangdong Provincial Hospital of Traditional Chinese Medicine (GPHTCM), 
      Guangzhou, China. Electronic address: zhengguangjuan@163.com.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20151125
PL  - Netherlands
TA  - Exp Mol Pathol
JT  - Experimental and molecular pathology
JID - 0370711
RN  - 0 (Fixatives)
RN  - 1HG84L3525 (Formaldehyde)
RN  - 8002-74-2 (Paraffin)
RN  - EC 2.7.10.1 (Receptor, ErbB-2)
SB  - IM
MH  - Adult
MH  - Aged
MH  - Aged, 80 and over
MH  - Breast Neoplasms/diagnosis/*genetics/metabolism
MH  - Cell Line, Tumor
MH  - Female
MH  - Fixatives/chemistry
MH  - Formaldehyde/chemistry
MH  - Gene Amplification
MH  - Humans
MH  - Immunohistochemistry
MH  - In Situ Hybridization, Fluorescence
MH  - Middle Aged
MH  - Paraffin
MH  - Polymerase Chain Reaction/*methods
MH  - Receptor, ErbB-2/*genetics/metabolism
MH  - Reproducibility of Results
MH  - Sensitivity and Specificity
MH  - Stomach Neoplasms/diagnosis/*genetics/metabolism
MH  - Tissue Embedding/methods
MH  - Tissue Fixation/methods
OTO - NOTNLM
OT  - Breast cancer
OT  - Droplet digital PCR
OT  - Gastric cancer
OT  - HER2 amplification
EDAT- 2015/12/03 06:00
MHDA- 2016/09/07 06:00
CRDT- 2015/12/03 06:00
PHST- 2015/11/10 00:00 [received]
PHST- 2015/11/24 00:00 [accepted]
PHST- 2015/12/03 06:00 [entrez]
PHST- 2015/12/03 06:00 [pubmed]
PHST- 2016/09/07 06:00 [medline]
AID - S0014-4800(15)00238-5 [pii]
AID - 10.1016/j.yexmp.2015.11.027 [doi]
PST - ppublish
SO  - Exp Mol Pathol. 2016 Apr;100(2):287-93. doi: 10.1016/j.yexmp.2015.11.027. Epub 
      2015 Nov 25.