1O - Detection of ALK rearranged non-small cell lung carcinomas by immunohistochemistry: comparison of different antibodies (D5F3 and 5A4), detection sys...
|Date||27 March 2014|
|Session||Proffered Papers 2 - Tumour biology and translational research|
|Topics|| Lung and other Thoracic Tumours
|Citation||Journal of Thoracic Oncology (2014) 9 (Supplement 9): S7-S52. 10.1097/JTO.0000000000000131|
S. Savic1, J. Diebold2, A. Zimmermann3, W. Jochum4, B. Baschiera1, R. Epper1, S. Grieshaber1, L. Tornillo1, K. Kerr5, L. Bubendorf1
Immunohistochemistry (IHC) has become a promising method for prescreening predictive ALK-rearrangements in non-small cell lung carcinomas (NSCLC). Various ALK antibodies, enhanced detection systems and automated immunostainers are available. We therefore aimed to compare the performance of: 1. the D5F3 (Cell Signaling, Ventana) and the 5A4 (Novocastra, Leica) monoclonal antibodies (mAb) and 2. the 5A4 mAb on two different automated immunostainers (BenchMark, Ventana and BOND-MAX, Leica).
Methods: Sixty-five formalin-fixed and paraffin-embedded non-squamous NSCLC with available ALK FISH results were retrospectively analysed. The specimens were enriched for ALK FISH-positive NSCLC (24/65, 36.9%). IHC was performed with the OptiView Amplification Kit (Ventana) on BenchMark using the D5F3 and the 5A4 mAb, respectively. Additionally IHC with the 5A4 mAb and the Bond Polymer Refine Detection Kit (Leica) on Bond-MAX was performed. For D5F3 any strong IHC-staining (3+) was regarded as a positive result, as suggested by the manufacturer. For 5A4 any IHC-staining (1+, 2+, 3+) was regarded as a positive result. The IHC results of all three protocols were compared to the FISH results. In case of a discrepancy the IHC-stained slide and the FISH signals were reviewed.