PMID- 17122642
OWN - NLM
STAT- MEDLINE
DCOM- 20070123
LR  - 20191210
IS  - 1541-2016 (Print)
IS  - 1533-4058 (Linking)
VI  - 14
IP  - 4
DP  - 2006 Dec
TI  - Automation of manual components and image quantification of direct dual label 
      fluorescence in situ hybridization (FISH) for HER2 gene amplification: A 
      feasibility study.
PG  - 436-40
AB  - Determination of HER2 status by fluorescence in situ hybridization (FISH) in 
      breast carcinoma correlates well with response to targeted therapy and prognosis. 
      However, manual time consuming methods and quantification aspects of the 
      procedure may be challenging for some laboratories. We examined the feasibility 
      of automating these components of the FISH assay using a tissue microarray 
      (TMA-118 clinically annotated cases) and a series of 41 whole sections. An in 
      situ hybridization automated staining workstation was used to automate a 
      programmed overnight start, on line baking, deparaffinization, cell conditioning, 
      protease digestion, and prehybridization buffer washing. Dual label probe/target 
      codenaturation/hybridization and stringency washing were done off line. The HER2 
      and CEP17 spot counts were quantified, and the HER2/CEP17 ratio calculated, via 
      an imaging workstation. Results were benchmarked against manual counts for whole 
      sections, and bright field in situ hybridization [silver in situ hybridization 
      (SISH)] for the TMA. Automated FISH results using whole sections correlated well 
      with manual results: HER2/CEP17 ratio correlation coefficient r = 0.9154, r = 
      0.8380, P < 0.0001. Correlation between automated and manual TMA FISH results was 
      also excellent, and disease-free survival was significantly shorter (P < 0.001) 
      for the HER2 amplified cases. Automation of the laborious manual prehybridization 
      and image quantification components of FISH using directly labeled probes is 
      feasible. Operational gains and enhanced consistency are inherent in this 
      automated approach to HER2 clinical FISH testing.
FAU - Tubbs, Raymond R
AU  - Tubbs RR
AD  - Department of Anatomic and Clinical Pathology, The Cleveland Clinic Foundation 
      and the Cleveland Clinic Lerner College of Medicine of Case Western Reserve 
      University, Cleveland, OH 44195, USA. tubbsr@ccf.org
FAU - Pettay, James D
AU  - Pettay JD
FAU - Swain, Eric
AU  - Swain E
FAU - Roche, Patrick C
AU  - Roche PC
FAU - Powell, William
AU  - Powell W
FAU - Hicks, David G
AU  - Hicks DG
FAU - Grogan, Thomas
AU  - Grogan T
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
PL  - United States
TA  - Appl Immunohistochem Mol Morphol
JT  - Applied immunohistochemistry & molecular morphology : AIMM
JID - 100888796
SB  - IM
MH  - Automation
MH  - Breast Neoplasms/*metabolism
MH  - Carcinoma, Ductal, Breast/*metabolism
MH  - Feasibility Studies
MH  - Female
MH  - *Gene Amplification
MH  - *Genes, erbB-2
MH  - Humans
MH  - In Situ Hybridization, Fluorescence/methods
MH  - Signal Processing, Computer-Assisted
MH  - Tissue Array Analysis
EDAT- 2006/11/24 09:00
MHDA- 2007/01/24 09:00
CRDT- 2006/11/24 09:00
PHST- 2006/11/24 09:00 [pubmed]
PHST- 2007/01/24 09:00 [medline]
PHST- 2006/11/24 09:00 [entrez]
AID - 00129039-200612000-00012 [pii]
AID - 10.1097/01.pai.0000213101.26193.f1 [doi]
PST - ppublish
SO  - Appl Immunohistochem Mol Morphol. 2006 Dec;14(4):436-40. doi: 
      10.1097/01.pai.0000213101.26193.f1.