Correlation of ROS1 (D4D6) Immunohistochemistry with ROS1 Fluorescence In Situ Hybridization Assay in a Contemporary Cohort of Pulmonary Adenocarcinomas

Shivani Sharma; Sourav K Mishra; Mohit Bhardwaj; Shilpy Jha; Matthew Geller; Aditi Dewan; Ekta Jain; Mallika Dixit; Deepika Jain; Gauri Munjal; Shivmurti Kumar; Sambit K Mohanty

doi:10.1055/s-0042-1750187

Correlation of ROS1 (D4D6) Immunohistochemistry with ROS1 Fluorescence In Situ Hybridization Assay in a Contemporary Cohort of Pulmonary Adenocarcinomas

South Asian J Cancer. 2022 Jun 25;11(3):249-255. doi: 10.1055/s-0042-1750187. eCollection 2022 Jul.

Authors

Affiliations

¹ Department of Pathology and Laboratory Medicine, CORE Diagnostics, Gurgaon, Haryana, India.
² Department of Medical Oncology, SUM Hospital, Bhubaneswar, Odisha, India.
³ Department of Pathology and Laboratory Medicine, Advanced Medical Research Institute, Bhubaneswar, Odisha, India.
⁴ Department of Pathology and Laboratory Medicine, Washington County Pathologists, PC Hillsboro, Oregon, United States.

Abstract

Sambit K. MohantyObjective Repressor of Silencing ( ROS1 ) gene rearrangement in the lung adenocarcinomas is one of the targetable mutually exclusive genomic alteration. Fluorescence in situ hybridization (FISH), immunohistochemistry (IHC), next-generation sequencing, and reverse transcriptase polymerase chain reaction assays are generally used to detect ROS1 gene alterations. We evaluated the correlation between ROS1 IHC and FISH analysis considering FISH as the gold standard method to determine the utility of IHC as a screening method for lung adenocarcinoma. Materials and Methods A total of 374 advanced pulmonary adenocarcinoma patients were analyzed for ROS1 IHC on Ventana Benchmark XT platform using D4D6 rabbit monoclonal antibody. FISH assay was performed in parallel in all these cases using the Vysis ROS1 Break Apart FISH probe. Statistical Analysis The sensitivity, specificity, positive and negative likelihood ratios, positive and negative predictive values, and accuracy were evaluated. Results A total of 17 tumors were positive either by IHC or FISH analysis or both (true positive). Four tumors were positive by IHC (H-score range: 120-270), while negative on FISH analysis (false positive by IHC). One tumor was IHC negative, but positive by FISH analysis (false negative). The sensitivity, specificity, positive likelihood ratio, negative likelihood ratio, positive predictive value, negative predictive value, and accuracy were 94.4% (confidence interval [CI]: 72.71-99.86%), 63.6% (CI: 30.79-89.07%), 2.6 (CI: 1.18-5.72), 0.09 (CI: 0.01-0.62), 80.95% (CI: 65.86-90.35%), 87.5% (CI: 49.74-98.02%), and 82.76%, respectively. Conclusion ROS1 IHC has high sensitivity at a cost of lower specificity for the detection of ROS1 gene rearrangement. All IHC positive cases should undergo a confirmatory FISH test as this testing algorithm stands as a reliable and economic tool to screen ROS1 rearrangement in lung adenocarcinomas.

Keywords: D4D6 clone; FISH; ROS1; immunohistochemistry; pulmonary adenocarcinoma.

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