title: A multicenter blinded study evaluating EGFR and KRAS mutation testing methods in the clinical non-small cell lung cancer setting--IFCT/ERMETIC2 Project Part 1: Comparison of testing methods in 20 French molecular genetic National Cancer Institute platforms.
abstract: Epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitors have limited use as first-line treatment for mutated EGFR metastatic non-small cell lung cancer. The French National Cancer Institute has installed molecular genetics platforms implementing EGFR and KRAS testing. However, there is considerable uncertainty as to which detection methods should be applied for routine diagnosis. This study aimed to compare the EGFR and KRAS genotyping methods developed by the IFCT/ERMETIC2 network platforms in two blind panels: 25 samples of serial dilutions of cell line DNA (20 centers) and 74 FFPE lung tumor samples (10 centers). The best threshold of mutation detection on cell lines was obtained using allele-specific amplification-based technologies. Nonamplifiable tissue samples were significantly less common when using alternative testing versus direct sequencing [15%; 95% confidence interval (CI), 14%-16% versus 40%; 95% CI, 39%-42%; P < 0.001]. Mutated cases increased from 42% (95% CI, 31%-54%) to 53% (95% CI, 41%-64%), with three supplementary EGFR mutations (p.G179A at exon 18 and p.L858R and p.L861Q at exon 21) and five supplementary KRAS mutations, when using alternative testing instead of direct sequencing. False-positive results were observed when using a PCR-based sizing assay, high-resolution melting, or pyrosequencing. Concordance analysis returned good kappa test scores for EGFR exon 19 and KRAS analysis when comparing sequencing with alternative methods and revealed no difference between alternative techniques themselves.
authors.affiliation: Department of Molecular Biology, Strasbourg University Hospital, EA 3430, Strasbourg University, Strasbourg, France; the Francophone Intergroup of Thoracic Oncology, Paris, France. Electronic address: michele.faller@chru-strasbourg.fr.
authors.forename: Michèle
authors.initials: M
authors.lastname: Beau-Faller
authors.affiliation: Department of Molecular Biology, Georges Pompidou European Hospital, AP-HP, Paris, Université Paris Descartes, Paris, France.
authors.forename: Hélène
authors.initials: H
authors.lastname: Blons
authors.affiliation: Department of Biostatistics and Epidemiology, Institut Gustave-Roussy, Villejuif, France.
authors.forename: Caroline
authors.initials: C
authors.lastname: Domerg
authors.affiliation: Department of Biostatistics and Epidemiology, Institut Gustave-Roussy, Villejuif, France.
authors.forename: Dorota
authors.initials: D
authors.lastname: Gajda
authors.affiliation: Department of Genetics, Laboratory of Medical Genetics, CHU de Caen, Caen, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Richard
authors.affiliation: Oncology and Molecular Genetics Laboratory, Division of Biochemistry and Molecular Biology, Center for Disease Biology, CHRU de Lille, Lille Cedex, France.
authors.forename: Fabienne
authors.initials: F
authors.lastname: Escande
authors.affiliation: Molecular Biology Laboratory, CHU de Montpellier, Montpellier, France.
authors.forename: Jérôme
authors.initials: J
authors.lastname: Solassol
authors.affiliation: Biochemistry Laboratory, CHU de Nantes, Nantes, France.
authors.forename: Marc G
authors.initials: MG
authors.lastname: Denis
authors.affiliation: Molecular Biology Laboratory, CHU de Clermont-Ferrand, Clermont-Ferrand, France.
authors.forename: Anne
authors.initials: A
authors.lastname: Cayre
authors.affiliation: Laboratory of Oncology and Biological Transfer, CHU de Marseille, Marseille, France.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Nanni-Metellus
authors.affiliation: Medical Genetics and Functional Genomics, INSERM UMR S910, Marseille, France.
authors.forename: Sylviane
authors.initials: S
authors.lastname: Olschwang
authors.affiliation: Molecular Biology Unit, Centre GF Leclerc, Dijon, France.
authors.forename: Sarab
authors.initials: S
authors.lastname: Lizard
authors.affiliation: Molecular Biology Laboratory, CHU de Dijon, Dijon, France.
authors.forename: Fabienne
authors.initials: F
authors.lastname: Piard
authors.affiliation: Univ Franche-Comte, EA 3181, FED4234, and CHU Besancon, Besancon, France.
authors.forename: Jean-Luc
authors.initials: JL
authors.lastname: Pretet
authors.affiliation: UM Biochemistry and Cancer Biotherapy, Division of Biology, CHU Grenoble, Grenoble Cedex 09, France.
authors.forename: Florence
authors.initials: F
authors.lastname: de Fraipont
authors.affiliation: Oncogenetics Laboratory, Plateforme HUVEGEN, Institut Curie-Hôpital René Huguenin, Saint-Cloud, France.
authors.forename: Ivan
authors.initials: I
authors.lastname: Bièche
authors.affiliation: Pharmacology Unit, Department of Tumor Biology, Institut Curie, Paris, France.
authors.forename: Patricia
authors.initials: P
authors.lastname: de Cremoux
authors.affiliation: Unit of Lung Molecular Pathology of the Hospital Platform of Cancer Molecular Genetics Midi-Pyrénées, Pathological Anatomy Service, CHU Toulouse Rangueil, Toulouse, France.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Rouquette
authors.affiliation: Department of Anatomy and Pathological Cytology, Hospices Civils de Lyon, Hôpital Edouard Herriot, Lyon, France.
authors.forename: Pierre-Paul
authors.initials: PP
authors.lastname: Bringuier
authors.affiliation: Laboratory of Somatic Cancer Genetics, CHU de Rennes, Rennes, France.
authors.forename: Jean
authors.initials: J
authors.lastname: Mosser
authors.affiliation: Molecular Biology Laboratory, Hôpital de Hautepierre, Strasbourg, France.
authors.forename: Michèle
authors.initials: M
authors.lastname: Legrain
authors.affiliation: Molecular Biology Laboratory, Hôpital de Hautepierre, Strasbourg, France.
authors.forename: Anne-Claire
authors.initials: AC
authors.lastname: Voegeli
authors.affiliation: Translational Research Laboratory, Institut Gustave Roussy, Villejuif Cedex, France.
authors.forename: Patrick
authors.initials: P
authors.lastname: Saulnier
authors.affiliation: the Francophone Intergroup of Thoracic Oncology, Paris, France.
authors.forename: Franck
authors.initials: F
authors.lastname: Morin
authors.affiliation: the Francophone Intergroup of Thoracic Oncology, Paris, France; Department of Molecular Biology, Georges Pompidou European Hospital, AP-HP, Paris, Université Paris Descartes, Paris, France.
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Pignon
authors.affiliation: the Francophone Intergroup of Thoracic Oncology, Paris, France; UMR 1086 INSERM and the Department of Pneumlogy and Thoracic Oncology, Caen University Hospital, Caen, France.
authors.forename: Gérard
authors.initials: G
authors.lastname: Zalcman
authors.affiliation: the Francophone Intergroup of Thoracic Oncology, Paris, France; Pneumology Service, Assistance Publique Hôpitaux de Paris, Hôpital Tenon, GRC-04 Theranoscan, Université Paris VI, Paris Cedex 20, France.
authors.forename: Jacques
authors.initials: J
authors.lastname: Cadranel
mesh_terms: D000230:Adenocarcinoma / Q000175:diagnosis* / Q000188:drug therapy / Q000235:genetics; D000077192:Adenocarcinoma of Lung; D000970:Antineoplastic Agents / Q000627:therapeutic use; D001483:Base Sequence; D002289:Carcinoma, Non-Small-Cell Lung / Q000175:diagnosis* / Q000188:drug therapy / Q000235:genetics; D045744:Cell Line, Tumor; D066246:ErbB Receptors / Q000037:antagonists & inhibitors / Q000235:genetics*; D000069347:Erlotinib Hydrochloride; D005189:False Positive Reactions; D000077156:Gefitinib; D005838:Genotype; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods; D006801:Humans; D008175:Lung Neoplasms / Q000175:diagnosis* / Q000188:drug therapy / Q000235:genetics; D025202:Molecular Diagnostic Techniques / Q000379:methods; D021141:Nucleic Acid Amplification Techniques; D047428:Protein Kinase Inhibitors / Q000627:therapeutic use; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D016283:Proto-Oncogene Proteins p21(ras); D011799:Quinazolines / Q000627:therapeutic use; D017422:Sequence Analysis, DNA / Q000379:methods*; D018631:ras Proteins / Q000235:genetics*
chemical_list: D000970:Antineoplastic Agents; C117307:KRAS protein, human; D047428:Protein Kinase Inhibitors; D011518:Proto-Oncogene Proteins; D011799:Quinazolines; D000069347:Erlotinib Hydrochloride; C512478:EGFR protein, human; D066246:ErbB Receptors; D016283:Proto-Oncogene Proteins p21(ras); D018631:ras Proteins; D000077156:Gefitinib