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*Breast Cancer
Journal of Clinical Pathology 2005;58:710-714
© 2005 BMJ Publishing Group Ltd & Association of Clinical Pathologists


ORIGINAL ARTICLE

HER2 amplification status in breast cancer: a comparison between immunohistochemical staining and fluorescence in situ hybridisation using manual and automated quantitative image analysis scoring techniques

C M Ellis1, M J Dyson1, T J Stephenson2, E L Maltby1

1 Department of Cytogenetics, Sheffield Genetics Services, Sheffield Children’s NHS Trust, Western Bank, Sheffield S10 2TH, UK
2 Department of Histopathology, Royal Hallamshire Hospital, Glossop Road, Sheffield S10 2SF, UK

Correspondence to:
Dr E L Maltby
Department of Cytogenetics, Sheffield Genetics Services, Sheffield Childrens NHS Trust, Western Bank, Sheffield S10 2TH, UK; Edna.Maltby{at}sheffch-tr.trent.nhs.uk Aims: To compare the results of breast cancer sections with HercepTestTM immunohistochemistry (IHC) scores ranging from 0 to 3+ with fluorescence in situ hybridisation (FISH) for HER2 amplification. The HER2 digital scoring application of the Micrometastasis Detection System (MDSTM) was used, together with manual scoring of FISH and HercepTest, to determine whether this system provides an accurate alternative.

Methods: Paraffin wax embedded sections were stained using HercepTest and analysed by eye and automated quantitative image analysis. FISH was performed using the PathVysionTM fluorescent probe and scored by eye and automated quantitative image analysis using MDS.

Results: Of 114 cases, 26% were amplified by FISH, whereas only 18% scored 3+; 32% of IHC 2+ cases were amplified by FISH, and one showed borderline amplification. Six percent of IHC negative cases (0 or 1+) were amplified by FISH, and one showed borderline amplification. Of IHC 3+ cases, 10% were non-amplified by FISH. Classification discrepancies were seen in 18% of HercepTest cases scored by eye and using the MDS system. MDS was consistent with visual FISH scoring and correctly differentiated most ambiguous visual IHC scores.

Conclusions: FISH provides a more accurate and consistent scoring system for determining HER2 amplification than HercepTest. The MDS system provides a reliable, consistent alternative to visual IHC and FISH scoring. IHC is still a valuable technique to aid in identification of isolated or heterogeneous tumour populations for subsequent FISH analysis, and a combined FISH and HercepTest approach to all breast cancer cases may be the most efficient strategy.


Abbreviations: DDW, double distilled water; FISH, fluorescence in situ hybridisation; IHC, immunohistochemistry; MDS, Micrometastasis Detection System

Keywords: FISH; HER2; breast cancer; Herceptin; immunohistochemistry




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