title: Fluorescence in situ hybridization, immunohistochemistry, and next-generation sequencing for detection of EML4-ALK rearrangement in lung cancer.
abstract: BACKGROUND
  The U.S. Food and Drug Administration-approved method for detecting EML4-ALK rearrangement is fluorescence in situ hybridization (FISH); however, data supporting the use of immunohistochemistry (IHC) for that purpose are accumulating. Previous studies that compared FISH and IHC considered FISH the gold standard, but none compared data with the results of next-generation sequencing (NGS) analysis.
  
  
  METHODS
  We studied FISH and IHC (D5F3 antibody) systematically for EML4-ALK rearrangement in 51 lung adenocarcinoma patients, followed by NGS in case of discordance.
  
  
  RESULTS
  Of 51 patients, 4 were positive with FISH (7.8%), and 8 were positive with IHC (15.7%). Three were positive with both. NGS confirmed that four of the five patients who were positive with IHC and negative with FISH were positive for ALK. Two were treated by crizotinib, with progression-free survival of 18 and 6 months. Considering NGS as the most accurate test, the sensitivity and specificity were 42.9% and 97.7%, respectively, for FISH and 100% and 97.7%, respectively, for IHC.
  
  
  CONCLUSIONS
  The FISH-based method of detecting EML4-ALK rearrangement in lung cancer may miss a significant number of patients who could benefit from targeted ALK therapy. Screening for EML4-ALK rearrangement by IHC should be strongly considered, and NGS is recommended in borderline cases. Two patients who were negative with FISH and positive with IHC were treated with crizotinib and responded to therapy.
authors.affiliation: Thoracic Cancer Research and Detection Center, Sheba Medical Center, Ramat Gan, Israel; Tel Aviv University, Tel Aviv, Israel; University of Colorado Cancer Center, Division of Medical Oncology, University of Colorado, Aurora, Colorado, USA; Oncotest, Teva Pharmaceutical Industries Ltd., Petah Tikva, Israel; Thoracic Cancer Unit, Davidoff Cancer Center, Rabin Medical Center, Petah Tikva, Israel; Foundation Medicine, Cambridge, Massachusetts, USA; The Institute of Oncology, Wolfson Medical Center, Holon, Israel.
authors.forename: Marina
authors.initials: M
authors.lastname: Pekar-Zlotin
authors.affiliation: Thoracic Cancer Research and Detection Center, Sheba Medical Center, Ramat Gan, Israel; Tel Aviv University, Tel Aviv, Israel; University of Colorado Cancer Center, Division of Medical Oncology, University of Colorado, Aurora, Colorado, USA; Oncotest, Teva Pharmaceutical Industries Ltd., Petah Tikva, Israel; Thoracic Cancer Unit, Davidoff Cancer Center, Rabin Medical Center, Petah Tikva, Israel; Foundation Medicine, Cambridge, Massachusetts, USA; The Institute of Oncology, Wolfson Medical Center, Holon, Israel.
authors.forename: Fred R
authors.initials: FR
authors.lastname: Hirsch
authors.affiliation: Thoracic Cancer Research and Detection Center, Sheba Medical Center, Ramat Gan, Israel; Tel Aviv University, Tel Aviv, Israel; University of Colorado Cancer Center, Division of Medical Oncology, University of Colorado, Aurora, Colorado, USA; Oncotest, Teva Pharmaceutical Industries Ltd., Petah Tikva, Israel; Thoracic Cancer Unit, Davidoff Cancer Center, Rabin Medical Center, Petah Tikva, Israel; Foundation Medicine, Cambridge, Massachusetts, USA; The Institute of Oncology, Wolfson Medical Center, Holon, Israel.
authors.forename: Lior
authors.initials: L
authors.lastname: Soussan-Gutman
authors.affiliation: Thoracic Cancer Research and Detection Center, Sheba Medical Center, Ramat Gan, Israel; Tel Aviv University, Tel Aviv, Israel; University of Colorado Cancer Center, Division of Medical Oncology, University of Colorado, Aurora, Colorado, USA; Oncotest, Teva Pharmaceutical Industries Ltd., Petah Tikva, Israel; Thoracic Cancer Unit, Davidoff Cancer Center, Rabin Medical Center, Petah Tikva, Israel; Foundation Medicine, Cambridge, Massachusetts, USA; The Institute of Oncology, Wolfson Medical Center, Holon, Israel.
authors.forename: Maya
authors.initials: M
authors.lastname: Ilouze
authors.affiliation: Thoracic Cancer Research and Detection Center, Sheba Medical Center, Ramat Gan, Israel; Tel Aviv University, Tel Aviv, Israel; University of Colorado Cancer Center, Division of Medical Oncology, University of Colorado, Aurora, Colorado, USA; Oncotest, Teva Pharmaceutical Industries Ltd., Petah Tikva, Israel; Thoracic Cancer Unit, Davidoff Cancer Center, Rabin Medical Center, Petah Tikva, Israel; Foundation Medicine, Cambridge, Massachusetts, USA; The Institute of Oncology, Wolfson Medical Center, Holon, Israel.
authors.forename: Addie
authors.initials: A
authors.lastname: Dvir
authors.affiliation: Thoracic Cancer Research and Detection Center, Sheba Medical Center, Ramat Gan, Israel; Tel Aviv University, Tel Aviv, Israel; University of Colorado Cancer Center, Division of Medical Oncology, University of Colorado, Aurora, Colorado, USA; Oncotest, Teva Pharmaceutical Industries Ltd., Petah Tikva, Israel; Thoracic Cancer Unit, Davidoff Cancer Center, Rabin Medical Center, Petah Tikva, Israel; Foundation Medicine, Cambridge, Massachusetts, USA; The Institute of Oncology, Wolfson Medical Center, Holon, Israel.
authors.forename: Theresa
authors.initials: T
authors.lastname: Boyle
authors.affiliation: Thoracic Cancer Research and Detection Center, Sheba Medical Center, Ramat Gan, Israel; Tel Aviv University, Tel Aviv, Israel; University of Colorado Cancer Center, Division of Medical Oncology, University of Colorado, Aurora, Colorado, USA; Oncotest, Teva Pharmaceutical Industries Ltd., Petah Tikva, Israel; Thoracic Cancer Unit, Davidoff Cancer Center, Rabin Medical Center, Petah Tikva, Israel; Foundation Medicine, Cambridge, Massachusetts, USA; The Institute of Oncology, Wolfson Medical Center, Holon, Israel.
authors.forename: Murry
authors.initials: M
authors.lastname: Wynes
authors.affiliation: Thoracic Cancer Research and Detection Center, Sheba Medical Center, Ramat Gan, Israel; Tel Aviv University, Tel Aviv, Israel; University of Colorado Cancer Center, Division of Medical Oncology, University of Colorado, Aurora, Colorado, USA; Oncotest, Teva Pharmaceutical Industries Ltd., Petah Tikva, Israel; Thoracic Cancer Unit, Davidoff Cancer Center, Rabin Medical Center, Petah Tikva, Israel; Foundation Medicine, Cambridge, Massachusetts, USA; The Institute of Oncology, Wolfson Medical Center, Holon, Israel.
authors.forename: Vincent A
authors.initials: VA
authors.lastname: Miller
authors.affiliation: Thoracic Cancer Research and Detection Center, Sheba Medical Center, Ramat Gan, Israel; Tel Aviv University, Tel Aviv, Israel; University of Colorado Cancer Center, Division of Medical Oncology, University of Colorado, Aurora, Colorado, USA; Oncotest, Teva Pharmaceutical Industries Ltd., Petah Tikva, Israel; Thoracic Cancer Unit, Davidoff Cancer Center, Rabin Medical Center, Petah Tikva, Israel; Foundation Medicine, Cambridge, Massachusetts, USA; The Institute of Oncology, Wolfson Medical Center, Holon, Israel.
authors.forename: Doron
authors.initials: D
authors.lastname: Lipson
authors.affiliation: Thoracic Cancer Research and Detection Center, Sheba Medical Center, Ramat Gan, Israel; Tel Aviv University, Tel Aviv, Israel; University of Colorado Cancer Center, Division of Medical Oncology, University of Colorado, Aurora, Colorado, USA; Oncotest, Teva Pharmaceutical Industries Ltd., Petah Tikva, Israel; Thoracic Cancer Unit, Davidoff Cancer Center, Rabin Medical Center, Petah Tikva, Israel; Foundation Medicine, Cambridge, Massachusetts, USA; The Institute of Oncology, Wolfson Medical Center, Holon, Israel.
authors.forename: Gary A
authors.initials: GA
authors.lastname: Palmer
authors.affiliation: Thoracic Cancer Research and Detection Center, Sheba Medical Center, Ramat Gan, Israel; Tel Aviv University, Tel Aviv, Israel; University of Colorado Cancer Center, Division of Medical Oncology, University of Colorado, Aurora, Colorado, USA; Oncotest, Teva Pharmaceutical Industries Ltd., Petah Tikva, Israel; Thoracic Cancer Unit, Davidoff Cancer Center, Rabin Medical Center, Petah Tikva, Israel; Foundation Medicine, Cambridge, Massachusetts, USA; The Institute of Oncology, Wolfson Medical Center, Holon, Israel.
authors.forename: Siraj M
authors.initials: SM
authors.lastname: Ali
authors.affiliation: Thoracic Cancer Research and Detection Center, Sheba Medical Center, Ramat Gan, Israel; Tel Aviv University, Tel Aviv, Israel; University of Colorado Cancer Center, Division of Medical Oncology, University of Colorado, Aurora, Colorado, USA; Oncotest, Teva Pharmaceutical Industries Ltd., Petah Tikva, Israel; Thoracic Cancer Unit, Davidoff Cancer Center, Rabin Medical Center, Petah Tikva, Israel; Foundation Medicine, Cambridge, Massachusetts, USA; The Institute of Oncology, Wolfson Medical Center, Holon, Israel.
authors.forename: Shlomi
authors.initials: S
authors.lastname: Dekel
authors.affiliation: Thoracic Cancer Research and Detection Center, Sheba Medical Center, Ramat Gan, Israel; Tel Aviv University, Tel Aviv, Israel; University of Colorado Cancer Center, Division of Medical Oncology, University of Colorado, Aurora, Colorado, USA; Oncotest, Teva Pharmaceutical Industries Ltd., Petah Tikva, Israel; Thoracic Cancer Unit, Davidoff Cancer Center, Rabin Medical Center, Petah Tikva, Israel; Foundation Medicine, Cambridge, Massachusetts, USA; The Institute of Oncology, Wolfson Medical Center, Holon, Israel.
authors.forename: Ronen
authors.initials: R
authors.lastname: Brenner
authors.affiliation: Thoracic Cancer Research and Detection Center, Sheba Medical Center, Ramat Gan, Israel; Tel Aviv University, Tel Aviv, Israel; University of Colorado Cancer Center, Division of Medical Oncology, University of Colorado, Aurora, Colorado, USA; Oncotest, Teva Pharmaceutical Industries Ltd., Petah Tikva, Israel; Thoracic Cancer Unit, Davidoff Cancer Center, Rabin Medical Center, Petah Tikva, Israel; Foundation Medicine, Cambridge, Massachusetts, USA; The Institute of Oncology, Wolfson Medical Center, Holon, Israel.
authors.forename: Paul A
authors.initials: PA
authors.lastname: Bunn
authors.affiliation: Thoracic Cancer Research and Detection Center, Sheba Medical Center, Ramat Gan, Israel; Tel Aviv University, Tel Aviv, Israel; University of Colorado Cancer Center, Division of Medical Oncology, University of Colorado, Aurora, Colorado, USA; Oncotest, Teva Pharmaceutical Industries Ltd., Petah Tikva, Israel; Thoracic Cancer Unit, Davidoff Cancer Center, Rabin Medical Center, Petah Tikva, Israel; Foundation Medicine, Cambridge, Massachusetts, USA; The Institute of Oncology, Wolfson Medical Center, Holon, Israel nirp@post.tau.ac.il.
authors.forename: Nir
authors.initials: N
authors.lastname: Peled
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics* / Q000378:metabolism* / Q000473:pathology; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000465:Algorithms; D003430:Cross-Sectional Studies; D018450:Disease Progression; D005260:Female; D015321:Gene Rearrangement; D006801:Humans; D007150:Immunohistochemistry; D017404:In Situ Hybridization, Fluorescence; D008175:Lung Neoplasms / Q000235:genetics* / Q000378:metabolism* / Q000473:pathology; D008297:Male; D008403:Mass Screening / Q000379:methods*; D008875:Middle Aged; D015514:Oncogene Proteins, Fusion / Q000235:genetics*; D012189:Retrospective Studies; D012680:Sensitivity and Specificity; D015996:Survival Rate
chemical_list: C522504:EML4-ALK fusion protein, human; D015514:Oncogene Proteins, Fusion
keywords: EML4-ALK; Fluorescence in situ hybridization; Immunohistochemistry; Next-generation sequencing; Non-small cell lung cancer