Correlation of ROS1 Immunohistochemistry With ROS1 Fusion Status Determined by Fluorescence In Situ Hybridization
Richard S P Huang1, Derek Smith1, Catherine H Le1
1From Roche Molecular Solutions, Pleasanton, California (Dr Huang, Mr Smith, Dr Le, Mr Liu, Dr Ordinario, Dr Manohar, Mr Lee, Mr Rajamani, Mr Truong, Dr Jing Li, Ms Choi, Dr Jingchuan Li, Dr Pati, Dr Hanlon Newell, Mr Pate, and Dr Menzl); Institute of Pathology, University Hospital Basel, Basel, Switzerland (Dr Bubendorf); Institute for Pathology, University Hospital, Cologne, Germany (Dr Buettner); the Department of Pathology, Aberdeen University Medical School & Aberdeen Royal Infirmary, Foresterhill, Aberdeen, United Kingdom (Dr Kerr); Laboratorio de Dianas Terapeuticas, HM Hospitales, Spain (Dr Lopez-Rios); Center of Predictive Molecular Medicine, CeSIMeT, University of Chieti-Pescara, Italy (Dr Marchetti); Pfizer Oncology, International Developed Markets, Berlin, Germany (Dr Marondel); the Department of Histopathology, Royal Brompton and Harefield Hospitals, and National Heart and Lung Division, Imperial College, London, United Kingdom (Dr Nicholson); the Pathology Department, Istanbul University, Cerrahpassa Medical Faculty, Istanbul, Turkey (Dr Öz); CORE, Antwerp University, and the Department of Pathology, Antwerp University Hospital, Antwerp, Belgium (Dr Pauwels); the Department of Biopathology, Centre Jean Perrin & INSERM U240 IMoST and Université Clermont Auvergne, Clermont-Ferrand, France (Dr Penault-Llorca); Operative Unit of Pathology, AUSL della Romagna, Hospital S. Maria delle Croci, Ravenna, Italy (Dr Rossi); and the Department of Pathology, VU University Medical Center, Amsterdam, the Netherlands (Dr Thunnissen).
Context.—:
The ability to determine ROS1 status has become mandatory for patients with lung adenocarcinoma, as many global authorities have approved crizotinib for patients with ROS1-positive lung adenocarcinoma.
Objective.—:
To present analytical correlation of the VENTANA ROS1 (SP384) Rabbit Monoclonal Primary Antibody (ROS1 [SP384] antibody) with ROS1 fluorescence in situ hybridization (FISH).
Design.—:
The immunohistochemistry (IHC) and FISH analytical comparison was assessed by using 122 non-small cell lung cancer samples that had both FISH (46 positive and 76 negative cases) and IHC staining results available. In addition, reverse transcription-polymerase chain reaction (RT-PCR) as well as DNA and RNA next-generation sequencing (NGS) were used to further examine the ROS1 status in cases that were discrepant between FISH and IHC, based on staining in the cytoplasm of 2+ or above in more than 30% of total tumor cells considered as IHC positive. Here, we define the consensus status as the most frequent result across the 5 different methods (IHC, FISH, RT-PCR, RNA NGS, and DNA NGS) we used to determine ROS1 status in these cases.
Results.—:
Of the IHC scoring methods examined, staining in the cytoplasm of 2+ or above in more than 30% of total tumor cells considered as IHC positive had the highest correlation with a FISH-positive status, reaching a positive percentage agreement of 97.8% and negative percentage agreement of 89.5%. A positive percentage agreement (100%) and negative percentage agreement (92.0%) was reached by comparing ROS1 (SP384) using a cutoff for staining in the cytoplasm of 2+ or above in more than 30% of total tumor cells to the consensus status.
Conclusions.—:
Herein, we present a standardized staining protocol for ROS1 (SP384) and data that support the high correlation between ROS1 status and ROS1 (SP384) antibody.
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