Romero, AtochaJantus-Lewintre, EloisaGarcía-Peláez, BeatrizRoyuela, AnaInsa, AmeliaCruz, PatriciaCollazo, AnaPérez Altozano, JavierVidal, Oscar JuanDiz, PilarCobo, ManuelHernández, BertaVázquez Estevez, SergioBenítez, GretelGuirado, MariaMajem, MargaritaBernabé, ReyesOrtega, Ana LauraBlasco, AnaBosch-Barrera, JoaquimJurado, Jose MGarcía González, JorgeViteri, SantiagoGarcia Giron, CarlosMassutí, BartomeuLopez Martín, AnaRodriguez-Festa, AlejandroCalabuig-Fariñas, SilviaMolina-Vila, Miguel ÁngelProvencio, Mariano2023-02-092023-02-092020-11-13http://hdl.handle.net/10668/16484Several platforms for noninvasive EGFR testing are currently used in the clinical setting with sensitivities ranging from 30% to 100%. Prospective studies evaluating agreement and sources for discordant results remain lacking. Herein, seven methodologies including two next-generation sequencing (NGS)-based methods, three high-sensitivity PCR-based platforms, and two FDA-approved methods were compared using 72 plasma samples, from EGFR-mutant non-small-cell lung cancer (NSCLC) patients progressing on a first-line tyrosine kinase inhibitor (TKI). NGS platforms as well as high-sensitivity PCR-based methodologies showed excellent agreement for EGFR-sensitizing mutations (K = 0.80-0.89) and substantial agreement for T790M testing (K = 0.77 and 0.68, respectively). Mutant allele frequencies (MAFs) obtained by different quantitative methods showed an excellent reproducibility (intraclass correlation coefficients 0.86-0.98). Among other technical factors, discordant calls mostly occurred at mutant allele frequencies (MAFs) ≤ 0.5%. Agreement significantly improved when discarding samples with MAF ≤ 0.5%. EGFR mutations were detected at significantly lower MAFs in patients with brain metastases, suggesting that these patients risk for a false-positive result. Our results support the use of liquid biopsies for noninvasive EGFR testing and highlight the need to systematically report MAFs.enAttribution 4.0 Internationalhttp://creativecommons.org/licenses/by/4.0/NGScirculating free DNAepidermal growth factor receptornon-small-cell lung cancerosimertinibtyrosine kinase inhibitorAdultAgedAged, 80 and overBiopsyCohort StudiesDNA Mutational AnalysisDNA, NeoplasmErbB ReceptorsExonsFemaleGene FrequencyHigh-Throughput Nucleotide SequencingHumansMaleMiddle AgedMutationPredictive Value of TestsSensitivity and SpecificitySequence Deletionresearch article33107189open access10.1002/1878-0261.128321878-0261PMC7782072https://febs.onlinelibrary.wiley.com/doi/pdfdirect/10.1002/1878-0261.12832https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7782072/pdf