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J Mol Diagn. 2014 Jan;16(1):45-55. doi: 10.1016/j.jmoldx.2013.07.009. Epub 2013 Oct 30.

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.

Author information

1
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.
2
Department of Molecular Biology, Georges Pompidou European Hospital, AP-HP, Paris, Université Paris Descartes, Paris, France.
3
Department of Biostatistics and Epidemiology, Institut Gustave-Roussy, Villejuif, France.
4
Department of Genetics, Laboratory of Medical Genetics, CHU de Caen, Caen, France.
5
Oncology and Molecular Genetics Laboratory, Division of Biochemistry and Molecular Biology, Center for Disease Biology, CHRU de Lille, Lille Cedex, France.
6
Molecular Biology Laboratory, CHU de Montpellier, Montpellier, France.
7
Biochemistry Laboratory, CHU de Nantes, Nantes, France.
8
Molecular Biology Laboratory, CHU de Clermont-Ferrand, Clermont-Ferrand, France.
9
Laboratory of Oncology and Biological Transfer, CHU de Marseille, Marseille, France.
10
Medical Genetics and Functional Genomics, INSERM UMR S910, Marseille, France.
11
Molecular Biology Unit, Centre GF Leclerc, Dijon, France.
12
Molecular Biology Laboratory, CHU de Dijon, Dijon, France.
13
Univ Franche-Comte, EA 3181, FED4234, and CHU Besancon, Besancon, France.
14
UM Biochemistry and Cancer Biotherapy, Division of Biology, CHU Grenoble, Grenoble Cedex 09, France.
15
Oncogenetics Laboratory, Plateforme HUVEGEN, Institut Curie-Hôpital René Huguenin, Saint-Cloud, France.
16
Pharmacology Unit, Department of Tumor Biology, Institut Curie, Paris, France.
17
Unit of Lung Molecular Pathology of the Hospital Platform of Cancer Molecular Genetics Midi-Pyrénées, Pathological Anatomy Service, CHU Toulouse Rangueil, Toulouse, France.
18
Department of Anatomy and Pathological Cytology, Hospices Civils de Lyon, Hôpital Edouard Herriot, Lyon, France.
19
Laboratory of Somatic Cancer Genetics, CHU de Rennes, Rennes, France.
20
Molecular Biology Laboratory, Hôpital de Hautepierre, Strasbourg, France.
21
Translational Research Laboratory, Institut Gustave Roussy, Villejuif Cedex, France.
22
the Francophone Intergroup of Thoracic Oncology, Paris, France.
23
the Francophone Intergroup of Thoracic Oncology, Paris, France; Department of Molecular Biology, Georges Pompidou European Hospital, AP-HP, Paris, Université Paris Descartes, Paris, France.
24
the Francophone Intergroup of Thoracic Oncology, Paris, France; UMR 1086 INSERM and the Department of Pneumlogy and Thoracic Oncology, Caen University Hospital, Caen, France.
25
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.

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.

PMID:
24183959
DOI:
10.1016/j.jmoldx.2013.07.009
[Indexed for MEDLINE]

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