SISH/CISH or qPCR as alternative techniques to FISH for determination of HER2 amplification status on breast tumors core needle biopsies: A multicenter experience based on 840 cases

Jocelyne Jacquemier, Frédérique Spyratos, Benjamin Esterni, Marie Joëlle Mozziconacci, Martine Antoine, Laurent Arnould, Sarab Lizard, Philippe Bertheau, Jacqueline Lehmann-Che, Cécile Blanc Fournier, Sophie Krieger, Frédéric Bibeau, Pierre Jean Lamy, Marie Pierre Chenard, Michèle Legrain, Jean Marc Guinebretière, Delphine Loussouarn, Gaëtan MacGrogan, Isabelle Hostein, Marie Christine MathieuLudovic Lacroix, Alexander Valent, Yves Marie Robin, Françoise Revillion, Magali Lacroix Triki, Aline Seaume, Anne Vincent Salomon, Patricia de Cremoux, Geneviève Portefaix, Luc Xerri, Sophie Vacher, Ivan Bièche, Frédérique Penault-Llorca

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    Abstract

    Background: Until now, FISH has been the gold standard technique to identify HER2 amplification status in ambiguous cases of breast cancer. Alternative techniques have been developed to increase the capacities of investigating HER2 amplification status. The aims of this multicenter study in a large series of breast cancer patients were to prospectively compare the level of performance of CISH, SISH, and qPCR alternative techniques on paraffin-embedded core biopsies with " gold standard FISH" for evaluation of HER2 amplification status.Methods: This study was performed on 840 cases scored by immunohistochemistry (IHC): 0=317 (38%), 1+=183 (22%), 2+=109 (13%), 3+=231 (27%). Each of the 15 French centers participating in the study analyzed 56 breast carcinoma cases diagnosed on fixed paraffin-embedded core biopsies. HER2 amplification status was determined by commercially available FISH used as the reference technique with determination of the HER2/CEN17 ratio or HER2 copy number status. The alternative techniques performed on the same cases were commercially available SISH or CISH and a common qPCR method especially designed for the study including a set of 10 primer pairs: 2 for HER2 (exons 8 and 26), 5 to evaluate chromosome 17 polysomy TAOK1, UTP6, MRM1, MKS1, SSTR2 and 3 for diploidy control TSN, LAP3 and ADAMTS16.Results: The concordance between IHC and FISH was 96% to 95% based on the HER2/CEN17 ratio (n=766) or HER2 copy number (n=840), respectively. The concordance of the alternative techniques with FISH was excellent: 97% and 98% for SISH (498 and 587 cases), 98% and 75% for CISH (108 and 204 cases) and 95% and 93% (699 and 773 cases) for qPCR based on the HER2/CEN17 ratio or HER2 copy number, respectively. Similarly, sensitivity ranged from 99% to 95% for SISH, 100% to 99% for CISH and 89% to 80% for qPCR. The concordance with FISH (ratio) in the 2+ cases was 89% for SISH, 100% for CISH and 93% for qPCR.Conclusion: These alternative techniques showed an excellent concordance with FISH in core biopsies allowing their use in routine clinical practice. This newly designed qPCR on paraffin-embedded core biopsies deserves special attention, as it is reliable, easy to perform and less expensive than ISH tests.

    Original languageEnglish
    Article number351
    JournalBMC Cancer
    Volume13
    DOIs
    Publication statusPublished - 22 Jul 2013

    Keywords

    • CISH
    • FISH
    • HER2
    • Multicenter analysis
    • SISH
    • qPCR

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