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Journal of Histochemistry and Cytochemistry
Volume 52 (4): 501-507, 2004
Copyright ©The Histochemical Society, Inc.

The Use of TMA for Interlaboratory Validation of FISH Testing for Detection of HER2 Gene Amplification in Breast Cancer

Leslie K. Diaz, Rohit Gupta, Noman Kidwai, Nour Sneige and Elizabeth L. Wiley

Department of Pathology, Feinberg School of Medicine, Northwestern University, Chicago, Illinois (LKD,RG,NK,ELW), and Department of Pathology, University of Texas M.D. Anderson Cancer Center, Houston, Texas (NS)

Correspondence to: Leslie K. Diaz, MD, Northwestern Memorial Hospital, 251 E. Huron St., Feinberg 7-325, Chicago, IL 60611. E-mail: l-diaz{at}northwestern.edu

HER2 fluorescence in situ hybridization (FISH) testing for breast cancer is largely limited to academic centers and commercial laboratories. As testing demands increase, methods for rapid and cost-effective technical validation and quality assessment will be required. Tissue microarray (TMA), a technique for high-throughput biomarker evaluation, could help facilitate these needs. Our objective was to assess the usefulness of TMA technology for validation of HER2 FISH testing. Two TMA blocks containing paired cores from 41 breast cancers were constructed. HER2 FISH was performed in parallel at two institutions and the results compared. One institution, with considerable HER2 FISH experience, served as the reference laboratory. HER2 chromogenic in situ hybridization (CISH) and immunohistochemistry (IHC) were compared to the FISH results. For positive and negative results, the concordance rate between laboratories was 100%. Using kappa statistical analysis to determine interobserver agreement, HER2 to chromosome 17 gene copy ratios showed strong agreement between laboratories with kappa = 0.85 (perfect agreement = 1.0). Four cases displaying low-level amplification by CISH contained chromosome 17 polysomy and gene copy ratios of <2.0 by FISH. Good concordance was observed between HER2 IHC and in situ hybridization testing. TMA is a robust and effective method for the technical validation of HER2 FISH testing and should be considered for use by quality assessment programs. (J Histochem Cytochem 52:501–507, 2004)

Key Words: breast cancer • HER2 • quality assurance • in situ hybridization


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