PMID- 23036330
OWN - NLM
STAT- MEDLINE
DCOM- 20130618
LR  - 20211203
IS  - 1095-9130 (Electronic)
IS  - 1046-2023 (Linking)
VI  - 59
IP  - 1
DP  - 2013 Jan
TI  - Droplet Digital PCR quantitation of HER2 expression in FFPE breast cancer 
      samples.
PG  - S20-3
LID - S1046-2023(12)00252-6 [pii]
LID - 10.1016/j.ymeth.2012.09.012 [doi]
AB  - The human epidermal growth factor receptor 2 (HER2, also known as erbB2) gene is 
      involved in signal transduction for cell growth and differentiation. It is a cell 
      surface receptor tyrosine kinase and a proto-oncogene. Overexpression of HER2 is 
      of clinical relevance in breast cancer due to its prognostic value correlating 
      elevated expression with worsening clinical outcome. At the same time, HER2 
      assessment is also of importance because successful anti-tumor treatment with 
      Herceptin(R) is strongly correlated with HER2 overexpression in the tumor 
      (approximately 30% of all breast tumors overexpress HER2). In a comprehensive 
      national study, Wolff et al. [1] state that "Approximately 20% of current HER2 
      testing may be inaccurate" which underscores the importance of developing more 
      accurate methods to determine HER2 status. Droplet Digital PCR (ddPCR) has the 
      potential to improve upon HER2 measurements due to its ability to quantitate DNA 
      and RNA targets with high precision and accuracy. Here we present a study which 
      investigates whether ddPCR can be used to assess HER2 transcript levels in 
      formalin-fixed paraffin embedded (FFPE) human breast tumors and whether these 
      ddPCR measurements agree with prior assessments of these same samples by 
      pathologists using immunohistochemistry (IHC) and in some cases fluorescence in 
      situ hybridization (FISH). We also determined the copy number of HER2 in these 
      samples as compared to the CEP17 reference gene. RESULTS: Clinical FFPE samples 
      were successfully studied using ddPCR and compared to results from standard FISH 
      and IHC methodology. The results demonstrate that ddPCR can rank order the 
      samples in complete agreement with the current standard methods and that ddPCR 
      has the added benefit of providing quantitative results, rather than relying on 
      the expert skill of a seasoned pathologist for determination.
CI  - Copyright (c) 2012 Elsevier Inc. All rights reserved.
FAU - Heredia, Nicholas J
AU  - Heredia NJ
AD  - Digital Biology Center, Bio-Rad Laboratories Inc., 7068 Koll Center Pkwy, Ste. 
      401, Pleasanton, CA 94566, USA.
FAU - Belgrader, Phillip
AU  - Belgrader P
FAU - Wang, Shenglong
AU  - Wang S
FAU - Koehler, Ryan
AU  - Koehler R
FAU - Regan, Jack
AU  - Regan J
FAU - Cosman, Angela M
AU  - Cosman AM
FAU - Saxonov, Serge
AU  - Saxonov S
FAU - Hindson, Benjamin
AU  - Hindson B
FAU - Tanner, Stephanie C
AU  - Tanner SC
FAU - Brown, Alexandra S
AU  - Brown AS
FAU - Karlin-Neumann, George
AU  - Karlin-Neumann G
LA  - eng
PT  - Comparative Study
PT  - Journal Article
DEP - 20121002
PL  - United States
TA  - Methods
JT  - Methods (San Diego, Calif.)
JID - 9426302
RN  - 0 (Fixatives)
RN  - 0 (MAS1 protein, human)
RN  - 0 (Proto-Oncogene Mas)
RN  - 0 (RNA, Messenger)
RN  - 1HG84L3525 (Formaldehyde)
RN  - EC 2.7.10.1 (ERBB2 protein, human)
RN  - EC 2.7.10.1 (Receptor, ErbB-2)
SB  - IM
MH  - Breast Neoplasms/*metabolism
MH  - Centromere/genetics
MH  - Female
MH  - Fixatives/chemistry
MH  - Formaldehyde/chemistry
MH  - Gene Dosage
MH  - *Gene Expression
MH  - Humans
MH  - Immunohistochemistry
MH  - In Situ Hybridization, Fluorescence
MH  - Molecular Diagnostic Techniques/*standards
MH  - Paraffin Embedding
MH  - Polymerase Chain Reaction/*standards
MH  - Proto-Oncogene Mas
MH  - RNA, Messenger/genetics/metabolism
MH  - Receptor, ErbB-2/*genetics/metabolism
MH  - Reference Standards
EDAT- 2012/10/06 06:00
MHDA- 2013/06/19 06:00
CRDT- 2012/10/06 06:00
PHST- 2012/10/06 06:00 [entrez]
PHST- 2012/10/06 06:00 [pubmed]
PHST- 2013/06/19 06:00 [medline]
AID - S1046-2023(12)00252-6 [pii]
AID - 10.1016/j.ymeth.2012.09.012 [doi]
PST - ppublish
SO  - Methods. 2013 Jan;59(1):S20-3. doi: 10.1016/j.ymeth.2012.09.012. Epub 2012 Oct 2.