HER2 status had been determined by HER2 fluorescence in situ hybridization or silver in situ hybridization (SISH) for cases that were equivocal by HER2 immunohistochemistry. In cases whose HER2 status was not determined, HER2 SISH assays were performed with INFORM HER2 DNA and chromosome 17 probes (Ventana Medical Systems) using an ultraView SISH Detection Kit (Ventana Medical Systems) as previously described [19 (link)]. At least 50 cells were evaluated for each case and HER2 status was determined according to the updated 2013 ASCO/CAP guidelines [18 (link)]. A HER2 copy number of 6.0 or higher per cell, or a HER2:CEP17 ratio of 2 or higher was defined as amplified. Cases with HER2/CEP17 ratios < 2 and HER2 copy numbers of 4 to 6 signals per cell were considered equivocal. HER2 copy numbers of < 4 signals per cell and HER2/CEP17 ratios < 2 were defined as non-amplified. In this study, HER2-equivocal cases were regarded as HER2–non-amplified for statistical analysis.
Inform her2 dna and chromosome 17 probes
Manufactured by Roche
Sourced in United States
The INFORM HER2 DNA and chromosome 17 probes are fluorescence in situ hybridization (FISH) probes designed for the detection of HER2 gene amplification and chromosome 17 copy number in formalin-fixed, paraffin-embedded (FFPE) human breast cancer tissue samples. The probes hybridize to specific DNA sequences on the HER2 gene and chromosome 17, allowing for the visualization and quantification of these genetic targets.
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4 protocols using inform her2 dna and chromosome 17 probes
1
HER2 Status Determination Protocol
2
HER2 Status Determination Protocols
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HER2 status was determined via HER2 fluorescence in situ hybridization or silver in situ hybridization (SISH) for cases with equivocal HER2 IHC. HER2 SISH was also performed in cases that showed discrepant results in HER2 status between primary and metastatic tumors. HER2 SISH assays were performed with INFORM HER2 DNA and Chromosome 17 probes (Ventana Medical Systems) using an ultraView SISH Detection Kit (Ventana Medical Systems) as previously described [17 (link)]. After scanning the whole section, at least 50 cells were evaluated in each case, and HER2 status was determined according to the 2013 ASCO/CAP guidelines [14 (link)]. In this study, HER2-equivocal cases were regarded as HER2-non-amplified for statistical analysis.
3
HER2 SISH Assay for Breast Cancer
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HER2 silver in situ hybridisation (SISH) assays were performed with INFORM HER2 DNA and Chromosome 17 probes (Ventana Medical Systems) using an ultraView SISH Detection Kit (Ventana Medical Systems) as previously described.27 (link) After scanning the whole section, at least 50 cells were evaluated in each case, and HER2 status was determined according to the updated 2013 ASCO/CAP guidelines.28 (link) HER2-equivocal cases were regarded as HER2 non-amplified.
4
Immunohistochemical Profiling of Biopsy Samples
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Formalin-fixed paraffin-embedded (FFPE) tissue sections, 4 μm thick, of
core-needle biopsy samples from all patients, and the surgical specimens of
the non-pCR group were subjected to immunohistochemical (IHC) staining for
estrogen receptor (6F11, 1:200, Novo Castra, Newcastle, UK), progesterone
receptor (PGR312, 1:200, Novo Castra), and HER2 (4B5, 1:8, Ventana, Tucson,
AZ, USA). IHC labeling was performed using an autostainer (Benchmark XT;
Ventana Medical Systems, AZ, USA) according to the manufacturer’s protocol.
Two pathologists reviewed these immune-labeled slides.
FFPE tissue sections, 4 μm thick, of samples scored as 2+ for HER2 were
subjected to automated silver in situ hybridization (SISH)
with INFORM HER2 DNA and chromosome 17 probes (Ventana Medical Systems, AZ,
USA), using an ultraView SISH Detection Kit (Ventana Medical Systems, AZ,
USA) according to the manufacturer’s instructions.
core-needle biopsy samples from all patients, and the surgical specimens of
the non-pCR group were subjected to immunohistochemical (IHC) staining for
estrogen receptor (6F11, 1:200, Novo Castra, Newcastle, UK), progesterone
receptor (PGR312, 1:200, Novo Castra), and HER2 (4B5, 1:8, Ventana, Tucson,
AZ, USA). IHC labeling was performed using an autostainer (Benchmark XT;
Ventana Medical Systems, AZ, USA) according to the manufacturer’s protocol.
Two pathologists reviewed these immune-labeled slides.
FFPE tissue sections, 4 μm thick, of samples scored as 2+ for HER2 were
subjected to automated silver in situ hybridization (SISH)
with INFORM HER2 DNA and chromosome 17 probes (Ventana Medical Systems, AZ,
USA), using an ultraView SISH Detection Kit (Ventana Medical Systems, AZ,
USA) according to the manufacturer’s instructions.
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