Consistency and reproducibility of next‐generation sequencing and other multigene mutational assays: A worldwide ring trial study on quantitative cytological molecular reference specimens

作者：Umberto Malapelle, Clara Mayo‐de‐las‐Casas, Miguel Ángel Molina‐Vila, Rafael Rosell, Spasenija Savic, Michel Bihl, Lukas Bubendorf, Manuel Salto‐Tellez, Dario de Biase, Giovanni Tallini, David H. Hwang, Lynette Marie Sholl, Rajyalakshmi Luthra, Birgit Weynand, Sara Vander Borght, Edoardo Missiaglia, Massimo Bongiovanni, Daniel Stieber, Philippe R. Vielh, Fernando Carlos Schmitt, Alessandra Rappa, Massimo C.P. Barberis, Francesco Pepe, Pasquale Pisapia, Nicola Serra, Elena Vigliar, Claudio Bellevicine, Matteo Fassan, Massimo Rugge, Carlos E. de Andrea, María D. Lozano, Fulvio Basolo, Gabriella Fontanini, Yuri E. Nikiforov, Suzanne Kamel‐Reid, Gilda da Cunha Santos, Marina N. Nikiforova, Sinchita Roy‐Chowdhuri, Giancarlo Troncone, The Molecular Cytopathology Meeting Group · 发表于：Cancer Cytopathology · 年份：2017 · DOI：10.1002/cncy.21868 · 被引用次数：89 · 研究领域：Lung Cancer Treatments and Mutations、Neuroblastoma Research and Treatments、Lung Cancer Research Studies

BACKGROUND: Molecular testing of cytological lung cancer specimens includes, beyond epidermal growth factor receptor (EGFR), emerging predictive/prognostic genomic biomarkers such as Kirsten rat sarcoma viral oncogene homolog (KRAS), neuroblastoma RAS viral [v-ras] oncogene homolog (NRAS), B-Raf proto-oncogene, serine/threonine kinase (BRAF), and phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit α (PIK3CA). Next-generation sequencing (NGS) and other multigene mutational assays are suitable for cytological specimens, including smears. However, the current literature reflects single-institution studies rather than multicenter experiences. METHODS: Quantitative cytological molecular reference slides were produced with cell lines designed to harbor concurrent mutations in the EGFR, KRAS, NRAS, BRAF, and PIK3CA genes at various allelic ratios, including low allele frequencies (AFs; 1%). This interlaboratory ring trial study included 14 institutions across the world that performed multigene mutational assays, from tissue extraction to data analysis, on these reference slides, with each laboratory using its own mutation analysis platform and methodology. RESULTS: All laboratories using NGS (n = 11) successfully detected the study's set of mutations with minimal variations in the means and standard errors of variant fractions at dilution points of 10% (P = .171) and 5% (P = .063) despite the use of different sequencing platforms (Illumina, Ion Torrent/Proton, and Roche...