HER2 FISH
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HER-2/neu (PathVysion ® ) FISH Assay Overview The HER-2 DNA Probe Kit (PathVysion ® Kit) is designed to detect amplification of the HER-2/neu gene via fluorescence in situ hybridization (FISH) in formalin-fixed paraffin-embedded (FFPE) human breast cancer tissue specimens. The PathVysion ® Kit (FDA approved) is indicated as an aid to predict disease-free and overall survival in patients with stage II, node-positive breast cancer who have been treated with adjuvant cyclophosphamide, doxorubicin and 5-fluorouracil (CAF) chemotherapy. The assay is also indicated as an aid in the assessment of patients for whom Herceptin ® (trastuzumab) treatment is being considered. Clinical Indications Breast Cancer. Clinical Utility Aids in predicting disease-free and overall survival in patients. Assesses candidates for Herceptin ® (trastuzumab) treatment. Methodology and Interpretation Fluorescence in situ hybridization (FISH). Signals for both HER-2/neu (red) and CEP17 (green) are scored. Amplification of HER- 2 gene is determined by the calculated the ratio HER-2/CEP17. However HER-2 amplification shows certain diversity due to co- amplification, intratumoral heterogeneity, and polysomy of chromosome 17: • Co-amplification happens when the HER-2 amplicon is large encompassing the centromere region as well. No evidence for HER-2/neu gene amplification by ratio cutoff would be seen as both HER-2 and the control CEP17 probe are amplified. However, FISH analysis is positive for HER-2 gene amplification. • Genetic heterogeneity (GH) of HER-2 is defined as presence of greater than 5% but less than 50% of infiltrating tumor cells with a HER-2/CEP17 ratio of greater than 2.2. At present, the clinical significance of GH in terms of the potential benefit from trastuzumab therapy is unclear. • The significance of increased HER-2 copy number associated with polysomy 17 with respect to response to trastuzumab is unclear. However, studies have shown a relationship between chromosome 17 polysomy and increased HER-2 protein expression. This raises the possibility that these individuals may respond to trastuzumab. Assay Specifications Reporting HER-2/neu Signals to Centromere 17 Ratio: >2.2 –– Amplified <1.8 –– Not Amplified 1.8-2.2 –– Equivocal Challenges in HER-2/neu testing addressed by FISH: Genetic Heterogeneity: Positive cells are >5% and <50% Equivocal Results: HER-2/CEP17 ratio is between 1.8 and 2.2 Specimen Requirements • 10% neutral buffered formalin-fixed paraffin-embedded (FFPE) tissue. • 3-5µm thick FFPE sections on positively coated slides. • Stored and transported at room temperature. Licensure CAP (Laboratory #: 7191582, AU-ID: 1434060), CLIA (Certificate #: 31D1038733), New Jersey (CLIS ID #: 0002299), New York State (PFI: 8192), Pennsylvania (031978), Florida (800018142), Maryland (1395) TAT 5-7 days CPT Codes 88368x2 201 Route 17 North • Rutherford • NJ 07070 • Office 201.528.9200 • Fax 201.528.9201 www.cancergenetics.com Breast Cancer © 2013 Cancer Genetics, Inc. All rights reserved. PathVysion ® is a registered trademark of Abbott Molecular.
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HER2 FISH
Transcript of HER2 FISH
HER-2/neu (PathVysion®) FISH AssayOverviewThe HER-2 DNA Probe Kit (PathVysion® Kit) is designed to detect amplification of the HER-2/neu gene via fluorescence in situ hybridization (FISH) in formalin-fixed paraffin-embedded (FFPE) human breast cancer tissue specimens. The PathVysion® Kit (FDA approved) is indicated as an aid to predict disease-free and overall survival in patients with stage II, node-positive breast cancer who have been treated with adjuvant cyclophosphamide, doxorubicin and 5-fluorouracil (CAF) chemotherapy. The assay is also indicated as an aid in the assessment of patients for whom Herceptin® (trastuzumab) treatment is being considered.
Clinical IndicationsBreast Cancer.
Clinical UtilityAids in predicting disease-free and overall survival in patients. Assesses candidates for Herceptin® (trastuzumab) treatment.
Methodology and InterpretationFluorescence in situ hybridization (FISH). Signals for both HER-2/neu (red) and CEP17 (green) are scored. Amplification of HER-2 gene is determined by the calculated the ratio HER-2/CEP17. However HER-2 amplification shows certain diversity due to co-amplification, intratumoral heterogeneity, and polysomy of chromosome 17:
• Co-amplification happens when the HER-2 amplicon is large encompassing the centromere region as well. No evidence forHER-2/neu gene amplification by ratio cutoff would be seen as both HER-2 and the control CEP17 probe are amplified. However, FISH analysis is positive for HER-2 gene amplification.
• Genetic heterogeneity (GH) of HER-2 is defined as presence of greater than 5% but less than 50% of infiltrating tumor cells with a HER-2/CEP17 ratio of greater than 2.2. At present, the clinical significance of GH in terms of the potential benefit from trastuzumab therapy is unclear.
• The significance of increased HER-2 copy number associated with polysomy 17 with respect to response to trastuzumab is unclear. However, studies have shown a relationship between chromosome 17 polysomy and increased HER-2 protein expression. This raises the possibility that these individuals may respond to trastuzumab.
Assay SpecificationsReportingHER-2/neu Signals to Centromere 17 Ratio: >2.2 –– Amplified <1.8 –– Not Amplified1.8-2.2 –– Equivocal
Challenges in HER-2/neu testing addressed by FISH:Genetic Heterogeneity: Positive cells are >5% and <50%Equivocal Results: HER-2/CEP17 ratio is between 1.8 and 2.2
Specimen Requirements• 10% neutral buffered formalin-fixed paraffin-embedded
(FFPE) tissue.• 3-5µm thick FFPE sections on positively coated slides.• Stored and transported at room temperature.
LicensureCAP (Laboratory #: 7191582, AU-ID: 1434060), CLIA (Certificate #: 31D1038733), New Jersey (CLIS ID #: 0002299), New York State (PFI: 8192), Pennsylvania (031978), Florida (800018142), Maryland (1395)
TAT5-7 days
CPT Codes88368x2
201 Route 17 North • Rutherford • NJ 07070 • Office 201.528.9200 • Fax 201.528.9201www.cancergenetics.com
Breast Cancer
© 2013 Cancer Genetics, Inc. All rights reserved.
PathVysion® is a registered trademark of Abbott Molecular.
201 Route 17 North • Rutherford • NJ 07070 • Office 201.528.9200 • Fax 201.528.9201www.cancergenetics.com
Patient Name: Accession Number:Sex: q Male q Female CGI ID No:
Date of Birth: Ordering Physician:Specimen: Client:Collected: Client Account No:Received: Client ID No:Reported: Client Address:
Clinical Hx: Telephone:
ERBB2 (HER2/NEU) FISH REPORT PATHVYSION® KIT
TOTAL # OF NUCLEI SCORED
60 RANGE OF ERBB2 FISH SIGNALS/NUCLEUS & HER2/CEP17 RATIO
Average ERBB2 signals/nucleus
18.37 HER2 gene copy<4 (N)HER2 gene copy 4-6(E)HER2 gene copy >6 (A)
Average cep17signals/nucleus
1.97 CEP 17 copy 2 (N,E,A)>2 in Polysomy of 17
Ratio of ERBB2/cep17 9.32 <1.8 (N) 1.8-2.2 (E) >2.2 (A)
Number of Readers 2
ISCN NOMENCULATURE: nuc ish(D17Z1x1.97),(ERBB2x18.37)[60]
Summary: POSITIVE FOR ERBB2 (HER2/neu) GENE AMPLIFICATION
FinalInterpretation:
The fluorescence in situ hybridization (FISH) analysis done on invasive breast carcinoma specimen (FFPE) showed AMPLIFIED signal patterns for ERBB2 (HER2/neu) both by HER2/CEP17 ratio and HER2 signals/nucleus criteria.
Ideogram: Probe Map:
End of ReportCEP 17 green/HER-2/neu red
17q11.1-q11.1 CEP 17alpha satelliteSpectrumGreen
17q11.2-q12 LSIHER-2/neuSpectrumOrange17
LSI HER-2
Centromere 17q11.2-q12 region Telomere
HER-2 gene
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© 2013 Cancer Genetics, Inc. All rights reserved.041013
