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Poster Display

1749 - Next generation sequencing identifies actionable mutations in EGFR-wild type and KRAS mutant non-small cell lung cancer patients


08 Oct 2016


Poster Display


Alexandra Voutsina


Annals of Oncology (2016) 27 (6): 416-454. 10.1093/annonc/mdw383


A. Voutsina1, A. Kalikaki1, A. Koutsopoulos2, M. Sfakianaki1, M. Trypaki1, E. Tsakalaki1, S. Agelaki3, V. Georgoulias4, D. Mavroudis5

Author affiliations

  • 1 Laboratory Of Translational Oncology, School of Medicine, University of Crete, 71003 - Heraklion/GR
  • 2 Pathology, School of Medicine, University of Crete, 71003 - Heraklion/GR
  • 3 Medical Oncology, Laboratory Of Translational Oncology, University Hospital of Heraklion, 71003 - Heraklion/GR
  • 4 Medical Oncology, Hellenic Oncology Research Group (HORG), 11471 - Athens/GR
  • 5 Medical Oncology, Laboratory Of Translational Oncology, University Hospital of Heraklion, 712 01 - Heraklion/GR


Abstract 1749


Identification of actionable mutations in patients' tumors is essential in guiding therapy. The aims of this study were: a) to validate mutation detection by next-generation sequencing (NGS) in a cohort of NSCLC patients and b) to identify molecular subgroups within EGFR wild-type and KRAS mutant NSCLCs


We used the Ion Torrent AmpliSeq Colon and Lung cancer panel to analyze formalin-fixed paraffin-embedded tumors from 76 NSCLC patients previously tested for EGFR mutations by Sanger Sequencing. The sensitivity of the method was assessed by using commercial reference FFPE standards with defined allelic frequencies. DNA was isolated from microdissected tumor tissue and sequencing was performed in the Ion PGM platform.


Tumors were sequenced to a median coverage of 650X. The sensitivity of mutation detection was estimated at 4% and for mutation reporting we have used a baseline prevalence of ≥5%. Precision of the method was demonstrated by analyzing four tumor specimens two times in different library preparations and runs. A complete concordance was observed between the previously defined Sanger genotypes and the corresponding variants detected using NGS. A single mutation was detected in 30 of 76 (39.5%) specimens, two in an additional 30 (39.5%) whereas mutations in more than two genes were detected in 11 (14.5%). The most frequently mutated genes were TP53 (41/76; 54%) and KRAS (23/76; 30%). Among KRAS mutated tumors, 3 (13%) carried STK11, 2 (8.7%) kinase inactivating BRAF mutations and one (4.4%) the BRAF V600E. Between the EGFR/KRAS wild type tumors, 5 (13%) had a mutation in the PI3K pathway, 3 (7.3%) carried mutations in MET, 2 (4.9%) had the BRAF V600E and 6 (14.6%) carried STK11 loss of function mutations


NGS can be used for molecular diagnostics in NSCLC and may detect additional mutated pathways that can be targeted using novel therapies

Clinical trial identification

Legal entity responsible for the study

Medical School, University of Crete


University of Crete


All authors have declared no conflicts of interest.

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