title: Clinical validation of a PCR assay for the detection of EGFR mutations in non-small-cell lung cancer: retrospective testing of specimens from the EURTAC trial.
abstract: The EURTAC trial demonstrated that the tyrosine kinase inhibitor (TKI) erlotinib was superior to chemotherapy as first-line therapy for advanced non-small cell lung cancers (NSCLC) that harbor EGFR activating mutations in a predominantly Caucasian population. Based on EURTAC and several Asian trials, anti-EGFR TKIs are standard of care for EGFR mutation-positive NSCLC. We sought to validate a rapid multiplex EGFR mutation assay as a companion diagnostic assay to select patients for this therapy. Samples from the EURTAC trial were prospectively screened for EGFR mutations using a combination of laboratory-developed tests (LDTs), and tested retrospectively with the cobas EGFR mutation test (EGFR PCR test). The EGFR PCR test results were compared to the original LDT results and to Sanger sequencing, using a subset of specimens from patients screened for the trial. Residual tissue was available from 487 (47%) of the 1044 patients screened for the trial. The EGFR PCR test showed high concordance with LDT results with a 96.3% overall agreement. The clinical outcome of patients who were EGFR-mutation detected by the EGFR PCR test was very similar to the entire EURTAC cohort. The concordance between the EGFR PCR test and Sanger sequencing was 90.6%. In 78.9% of the discordant samples, the EGFR PCR test result was confirmed by a sensitive deep sequencing assay. This retrospective study demonstrates the clinical utility of the EGFR PCR test in the accurate selection of patients for anti-EGFR TKI therapy. The EGFR PCR test demonstrated improved performance relative to Sanger sequencing.
authors.affiliation: Pangaea Biotech SL, Barcelona, Spain.
authors.forename: Susana
authors.initials: S
authors.lastname: Benlloch
authors.affiliation: Pangaea Biotech SL, Barcelona, Spain.
authors.forename: Maria Luisa
authors.initials: ML
authors.lastname: Botero
authors.affiliation: Pangaea Biotech SL, Barcelona, Spain.
authors.forename: Jordi
authors.initials: J
authors.lastname: Beltran-Alamillo
authors.affiliation: Pangaea Biotech SL, Barcelona, Spain.
authors.forename: Clara
authors.initials: C
authors.lastname: Mayo
authors.affiliation: Pangaea Biotech SL, Barcelona, Spain.
authors.forename: Ana
authors.initials: A
authors.lastname: Gimenez-Capitán
authors.affiliation: Medical Oncology Service-ICO, Hospital Germans Trias i Pujol, Badalona, Spain.
authors.forename: Itziar
authors.initials: I
authors.lastname: de Aguirre
authors.affiliation: Medical Oncology Service-ICO, Hospital Germans Trias i Pujol, Badalona, Spain.
authors.forename: Cristina
authors.initials: C
authors.lastname: Queralt
authors.affiliation: Medical Oncology Service-ICO, Hospital Germans Trias i Pujol, Badalona, Spain.
authors.forename: Jose Luis
authors.initials: JL
authors.lastname: Ramirez
authors.affiliation: Pangaea Biotech SL, Barcelona, Spain ; Pathology Department, Vall d'Hebron, University Hospital, Universidad Autónoma de Barcelona, Barcelona, Spain.
authors.forename: Santiago
authors.initials: S
authors.lastname: Ramón y Cajal
authors.affiliation: F. Hoffmann-La Roche, Basel, Switzerland.
authors.forename: Barbara
authors.initials: B
authors.lastname: Klughammer
authors.affiliation: F. Hoffmann-La Roche, Basel, Switzerland.
authors.forename: Mariette
authors.initials: M
authors.lastname: Schlegel
authors.affiliation: F. Hoffmann-La Roche, Basel, Switzerland.
authors.forename: Walter
authors.initials: W
authors.lastname: Bordogna
authors.affiliation: Genentech, South San Francisco, California, United States of America.
authors.forename: David
authors.initials: D
authors.lastname: Chen
authors.affiliation: Roche Molecular Systems, Pleasanton, California, United States of America.
authors.forename: Guili
authors.initials: G
authors.lastname: Zhang
authors.affiliation: Roche Molecular Systems, Pleasanton, California, United States of America ; Roche Molecular Systems, Pleasanton, California, United States of America.
authors.forename: Barbara
authors.initials: B
authors.lastname: Kovach
authors.affiliation: Roche Molecular Systems, Pleasanton, California, United States of America ; Roche Molecular Systems, Pleasanton, California, United States of America.
authors.forename: Felice
authors.initials: F
authors.lastname: Shieh
authors.affiliation: Roche Molecular Systems, Pleasanton, California, United States of America.
authors.forename: John F
authors.initials: JF
authors.lastname: Palma
authors.affiliation: Roche Molecular Systems, Pleasanton, California, United States of America.
authors.forename: Lin
authors.initials: L
authors.lastname: Wu
authors.affiliation: Roche Molecular Systems, Pleasanton, California, United States of America ; Roche Molecular Systems, Pleasanton, California, United States of America.
authors.forename: H Jeffrey
authors.initials: HJ
authors.lastname: Lawrence
authors.affiliation: Pangaea Biotech SL, Barcelona, Spain ; Medical Oncology Service-ICO, Hospital Germans Trias i Pujol, Badalona, Spain.
authors.forename: Miquel
authors.initials: M
authors.lastname: Taron
mesh_terms: D002289:Carcinoma, Non-Small-Cell Lung / Q000175:diagnosis* / Q000188:drug therapy / Q000235:genetics*; D066246:ErbB Receptors / Q000235:genetics*; D000069347:Erlotinib Hydrochloride; D005260:Female; D006801:Humans; D008175:Lung Neoplasms / Q000175:diagnosis* / Q000188:drug therapy / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D009154:Mutation / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods; D011446:Prospective Studies; D047428:Protein Kinase Inhibitors / Q000627:therapeutic use; D011799:Quinazolines / Q000627:therapeutic use; D012189:Retrospective Studies; D017422:Sequence Analysis, DNA / Q000379:methods
chemical_list: D047428:Protein Kinase Inhibitors; D011799:Quinazolines; D000069347:Erlotinib Hydrochloride; C512478:EGFR protein, human; D066246:ErbB Receptors