Malapelle Umberto, Mayo-de-Las-Casas Clara, Molina-Vila Miguel A, Rosell Rafael, Savic Spasenija, Bihl Michel, Bubendorf Lukas, Salto-Tellez Manuel, de Biase Dario, Tallini Giovanni, Hwang David H, Sholl Lynette M, Luthra Rajyalakshmi, Weynand Birgit, Vander Borght Sara, Missiaglia Edoardo, Bongiovanni Massimo, Stieber Daniel, Vielh Philippe, Schmitt Fernando, Rappa Alessandra, Barberis Massimo, Pepe Francesco, Pisapia Pasquale, Serra Nicola, Vigliar Elena, Bellevicine Claudio, Fassan Matteo, Rugge Massimo, de Andrea Carlos E, Lozano Maria D, Basolo Fulvio, Fontanini Gabriella, Nikiforov Yuri E, Kamel-Reid Suzanne, da Cunha Santos Gilda, Nikiforova Marina N, Roy-Chowdhuri Sinchita, Troncone Giancarlo
Department of Public Health, University of Naples Federico II, Naples, Italy.
Laboratory of Oncology, Pangaea Biotech, Barcelona, Spain.
Cancer Cytopathol. 2017 Aug;125(8):615-626. doi: 10.1002/cncy.21868. Epub 2017 May 5.
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.
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.
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). However, when mutations at a low AF of 1% were analyzed, the concordance of the NGS results was low, and this reflected the use of different thresholds for variant calling among the institutions. In contrast, laboratories using matrix-assisted laser desorption/ionization-time of flight (n = 2) showed lower concordance in terms of mutation detection and mutant AF quantification.
Quantitative molecular reference slides are a useful tool for monitoring the performance of different multigene mutational assays, and this could lead to better standardization of molecular cytopathology procedures. Cancer Cytopathol 2017;125:615-26. © 2017 American Cancer Society.
肺癌细胞学标本的分子检测，除了表皮生长因子受体（EGFR）外，还包括一些新出现的预测/预后基因组生物标志物，如 Kirsten 大鼠肉瘤病毒癌基因同源物（KRAS）、神经母细胞瘤 RAS 病毒[v-ras]癌基因同源物（NRAS）、B-Raf 原癌基因、丝氨酸/苏氨酸激酶（BRAF）以及磷脂酰肌醇-4,5-二磷酸 3-激酶催化亚基α（PIK3CA）。新一代测序（NGS）和其他多基因突变检测方法适用于细胞学标本，包括涂片。然而，目前的文献多为单机构研究，而非多中心经验。
使用设计为在 EGFR、KRAS、NRAS、BRAF 和 PIK3CA 基因中以各种等位基因比例（包括低等位基因频率（AFs；1%））同时存在突变的细胞系制作定量细胞学分子参考玻片。这项实验室间环形试验研究包括来自世界各地的 14 个机构，这些机构对这些参考玻片进行了从组织提取到数据分析的多基因突变检测，每个实验室使用自己的突变分析平台和方法。
所有使用 NGS 的实验室（n = 11）均成功检测到研究设定的突变，在 10%（P = 0.171）和 5%（P = 0.063）稀释点处，变异分数的均值和标准误差变化最小，尽管使用了不同的测序平台（Illumina、Ion Torrent/Proton 和 Roche）。然而，当分析 1%的低 AF 突变时，NGS 结果的一致性较低，这反映了各机构在变异调用时使用了不同的阈值。相比之下，使用基质辅助激光解吸/电离飞行时间质谱的实验室（n = 2）在突变检测和突变 AF 定量方面的一致性较低。
定量分子参考玻片是监测不同多基因突变检测性能的有用工具，这可能会使分子细胞病理学程序得到更好的标准化。《癌症细胞病理学》2017 年；125：615 - 26。©2017 美国癌症协会。
