PMID- 23358413
OWN - NLM
STAT- MEDLINE
DCOM- 20130805
LR  - 20220318
IS  - 1530-8561 (Electronic)
IS  - 0009-9147 (Linking)
VI  - 59
IP  - 6
DP  - 2013 Jun
TI  - Droplet digital PCR measurement of HER2 copy number alteration in formalin-fixed 
      paraffin-embedded breast carcinoma tissue.
PG  - 991-4
LID - 10.1373/clinchem.2012.197855 [doi]
AB  - BACKGROUND: Human epidermal growth factor receptor 2 (HER2) testing is routinely 
      performed by immunohistochemistry (IHC) and/or fluorescence in situ hybridization 
      (FISH) analyses for all new cases of invasive breast carcinoma. IHC is easier to 
      perform, but analysis can be subjective and variable. FISH offers better 
      diagnostic accuracy and added confidence, particularly when it is used to 
      supplement weak IHC signals, but it is more labor intensive and costly than IHC. 
      We examined the performance of droplet digital PCR (ddPCR) as a more precise and 
      less subjective alternative for quantifying HER2 DNA amplification. METHODS: 
      Thirty-nine cases of invasive breast carcinoma containing >/=30% tumor were 
      classified as positive or negative for HER2 by IHC, FISH, or both. DNA templates 
      for these cases were prepared from formalin-fixed paraffin-embedded (FFPE) 
      tissues to determine the HER2 copy number by ddPCR. ddPCR involved emulsifying 
      hydrolysis probe-based PCR reaction mixtures containing the ERBB2 [v-erb-b2 
      erythroblastic leukemia viral oncogene homolog 2, neuro/glioblastoma derived 
      oncogene homolog (avian); also known as HER2] gene and chromosome 17 centromere 
      assays into nanoliter-sized droplets for thermal cycling and analysis. RESULTS: 
      ddPCR distinguished, through differences in the level of HER2 amplification, the 
      10 HER2-positive samples from the 29 HER2-negative samples with 100% concordance 
      to HER2 status obtained by FISH and IHC analysis. ddPCR results agreed with the 
      FISH results for the 6 cases that were equivocal by IHC analyses, confirming 2 of 
      these samples as positive for HER2 and the other 4 as negative. CONCLUSIONS: 
      ddPCR can be used as a molecular-analysis tool to precisely measure copy number 
      alterations in FFPE samples of heterogeneous breast tumor tissue.
FAU - Belgrader, Phillip
AU  - Belgrader P
AD  - Digital Biology Center, Bio-Rad Laboratories, Pleasanton, CA 94566, USA. 
      phil_belgrader@bio-rad.com
FAU - Tanner, Stephanie C
AU  - Tanner SC
FAU - Regan, John F
AU  - Regan JF
FAU - Koehler, Ryan
AU  - Koehler R
FAU - Hindson, Benjamin J
AU  - Hindson BJ
FAU - Brown, Alexandra S
AU  - Brown AS
LA  - eng
GR  - R01EB010106/EB/NIBIB NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20130128
PL  - England
TA  - Clin Chem
JT  - Clinical chemistry
JID - 9421549
RN  - 1HG84L3525 (Formaldehyde)
RN  - EC 2.7.10.1 (Receptor, ErbB-2)
SB  - IM
MH  - Breast Neoplasms/*genetics
MH  - Computers
MH  - Female
MH  - Formaldehyde/chemistry
MH  - *Gene Dosage
MH  - Genetic Techniques/*standards
MH  - Humans
MH  - Paraffin Embedding
MH  - Polymerase Chain Reaction/*standards
MH  - Receptor, ErbB-2/*genetics
MH  - Reproducibility of Results
EDAT- 2013/01/30 06:00
MHDA- 2013/08/06 06:00
CRDT- 2013/01/30 06:00
PHST- 2013/01/30 06:00 [entrez]
PHST- 2013/01/30 06:00 [pubmed]
PHST- 2013/08/06 06:00 [medline]
AID - clinchem.2012.197855 [pii]
AID - 10.1373/clinchem.2012.197855 [doi]
PST - ppublish
SO  - Clin Chem. 2013 Jun;59(6):991-4. doi: 10.1373/clinchem.2012.197855. Epub 2013 Jan 
      28.