PMID- 11073807
OWN - NLM
STAT- MEDLINE
DCOM- 20001207
LR  - 20220408
IS  - 0002-9440 (Print)
IS  - 1525-2191 (Electronic)
IS  - 0002-9440 (Linking)
VI  - 157
IP  - 5
DP  - 2000 Nov
TI  - Chromogenic in situ hybridization: a practical alternative for fluorescence in 
      situ hybridization to detect HER-2/neu oncogene amplification in archival breast 
      cancer samples.
PG  - 1467-72
AB  - Determination of HER-2/neu oncogene amplification has become necessary for 
      selection of breast cancer patients for trastuzumab (Herceptin) therapy. 
      Fluorescence in situ hybridization (FISH) is currently regarded as a gold 
      standard method for detecting HER-2/neu amplification, but it is not very 
      practical for routine histopathological laboratories. We evaluated a new 
      modification of in situ hybridization, the chromogenic in situ hybridization 
      (CISH), which enables detection of HER-2/neu gene copies with conventional 
      peroxidase reaction. Archival formalin-fixed paraffin-embedded tumor tissue 
      sections were pretreated (by heating in a microwave oven and using enzyme 
      digestion) and hybridized with a digoxigenin-labeled DNA probe. The probe was 
      detected with anti-digoxigenin fluorescein, anti-fluorescein peroxidase, and 
      diaminobenzidine. Gene copies visualized by CISH could be easily distinguished 
      with a x40 objective in hematoxylin-stained tissue sections. HER-2/neu 
      amplification typically appeared as large peroxidase-positive intranuclear gene 
      copy clusters. CISH and FISH (according to Vysis, made from frozen pulverized 
      tumor samples) correlated well in a series of 157 breast cancers (kappa 
      coefficient, 0.81). The few different classifications were mostly because of 
      low-level amplifications by FISH that were negative by CISH and 
      immunohistochemistry with monoclonal antibody CB-11. We conclude that CISH, using 
      conventional bright-field microscopy in evaluation, is a useful alternative for 
      determination of HER-2/neu amplification in paraffin-embedded tumor samples, 
      especially for confirming the immunohistochemical staining results.
FAU - Tanner, M
AU  - Tanner M
AD  - Laboratory of Cancer Genetics, Institute of Medical Technology University and 
      University Hospital of Tampere, Tampere, Finland.
FAU - Gancberg, D
AU  - Gancberg D
FAU - Di Leo, A
AU  - Di Leo A
FAU - Larsimont, D
AU  - Larsimont D
FAU - Rouas, G
AU  - Rouas G
FAU - Piccart, M J
AU  - Piccart MJ
FAU - Isola, J
AU  - Isola J
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Am J Pathol
JT  - The American journal of pathology
JID - 0370502
RN  - 0 (Chromogenic Compounds)
SB  - IM
MH  - Archives
MH  - Breast Neoplasms/*genetics
MH  - *Chromogenic Compounds
MH  - Female
MH  - *Gene Amplification
MH  - Genes, erbB-2/*genetics
MH  - Humans
MH  - Immunohistochemistry/standards
MH  - In Situ Hybridization/*methods/standards
MH  - In Situ Hybridization, Fluorescence/standards
PMC - PMC1885742
EDAT- 2000/11/14 11:00
MHDA- 2001/02/28 10:01
PMCR- 2001/05/01
CRDT- 2000/11/14 11:00
PHST- 2000/11/14 11:00 [pubmed]
PHST- 2001/02/28 10:01 [medline]
PHST- 2000/11/14 11:00 [entrez]
PHST- 2001/05/01 00:00 [pmc-release]
AID - S0002-9440(10)64785-2 [pii]
AID - 2385 [pii]
AID - 10.1016/S0002-9440(10)64785-2 [doi]
PST - ppublish
SO  - Am J Pathol. 2000 Nov;157(5):1467-72. doi: 10.1016/S0002-9440(10)64785-2.