Non-Small-Cell Lung Cancer-Sensitive Detection of the p.Thr790Met EGFR Alteration by Preamplification before PNA-Mediated PCR Clamping and Pyrosequencing

• DOI: 10.3390/diagnostics10080527
• Publication Date: 2020-07-30T07:36:38Z
• Authors: Amandine Billaud; Veronique Verriele; Jonathan Dauvé; Louise-Marie Chev...; Alain Morel
• Keywords: Pyrosequencing; Peptide nucleic acid

Targeted therapies and, more precisely, EGFR tyrosine kinase inhibitors (TKIs) have been a major improvement in the therapeutic management of EGFR-mutated non-small-cell lung cancers (NSCLCs). Earlier administration these TKIs throughout tumor progression is imperative to improve patient outcomes. Consequently, studies focused on refining characterization biomarkers, especially concerning resistance mutation p.Thr790Met EGFR. Herein, we developed peptide nucleic acid (PNA)-mediated PCR clamping followed by pyrosequencing, favoring enrichment mutated fraction. A preamplification step was first added increase amplifiable DNA Throughout application our method extracted from FFPE samples 46 patients with NSCLC who had relapsed under first-generation TKI, evaluated sensitivity 93.3% and specificity 100%. All 19 were positive for NGS also found be protocol. The only discordant case sample no detected NGS, but which PNA. This protocol allows detection 0.5% will permit earlier an management.