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HER2 Testing in Breast Cancer

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8 Optimal Algorithm Figure 2. Evaluation of HER2 Gene Amplification by ISH Assay of the Invasive Component of a Breast Cancer Specimen Using a Single-signal (HER2 Gene) Assay (Single-probe ISH) HER2 testing (invasive component) by validated single-probe ISH assay Batch controls and on-slide controls show appropriate hybridization Average HER2 copy number ≥6.0 signals/cell a Concurrent IHC 3+ and/or Concurrent dual- probe ISH Group 1 c Average HER2 copy number <4.0 signals/cell a Concurrent IHC 2+ Perform dual-probe ISH for final result c NOTE. e final reported results assume that there is no apparent histopathologic discordance observed by the pathologist. a It is recommended that concomitant IHC review should become part of the interpretation of single- probe ISH results. e expert panel also preferentially recommends the use of dual-probe instead of single-probe ISH assays. b Using sections from the same tissue samples used for single-probe ISH, perform IHC (if not already performed) and/or dual-probe ISH. If IHC results are 2+ equivocal, it is recommended to also performdual probe ISH. c If initial assessment of dual-probe ISH suggestive of Groups 2, 3, or 4, follow the algorithm described in Figure3. Average HER2 copy number ≥4.0 and <6.0 signals/cell b Concurrent IHC 0, 1+ and/or Concurrent dual- probe ISH Group 5 c ISH positive ISH negative

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