RT Journal Article
SR Electronic
T1 Evaluation of four different molecular methods for EGFR mutation analysis in exon 18, 19 and 21
JF European Respiratory Journal
JO Eur Respir J
FD European Respiratory Society
SP p1958
VO 38
IS Suppl 55
A1 Anna Streubel
A1 Andreas Roth
A1 Susann Stephan-Falkenau
A1 Olfert Landt
A1 Cornelia Götz
A1 Thomas Mairinger
YR 2011
UL http://erj.ersjournals.com/content/38/Suppl_55/p1958.abstract
AB Aims: EGFR mutation status detection is essential for comprehensive NSCLC treatment Most Non Small Cell Lung Carcinomas are analyzed by histological examination of a biopsy obtained by bronchoscopy. Hence, only a small amount of tissue is available. These conditions require methods,suitable for the detection of very small amounts of mutated DNA. The aim of this study was to compare the sensitivity of mutation detection of four different molecular methods.Methods: 237 cases with NSCLC were included. All biopsies were formalin fixed and paraffin embedded. DNA was isolated after microdissection of tumor tissue from 20 μm slides, exon 18, 19 and 21 of the EGFR gene were investigated using pyrosequencing (PS) and BioFilm Chip Hybridization (Chip). Additionally, a LightCycler Assay was performed for Exon 19 (LC). All mutations were independently confirmed by Sanger sequencing.Results: We determined 197 cases without and 40 with mutations, 26 mutated in Exon 19. Chip: 24 samples, of which 4 mutated, could not be analyzed due to insufficient DNA amplification. Exon 19: 26 out of 26 detected correctly. PS: 39 of 40 mutations were detected. Exon19: 25 out of 26 detedcted correctly. LC (target only Exon 19): 25 of 26 were recognized correctly by this method.Discussion: A comparison of methods showed no significant difference in sensitivity. However, the Chipmethod failed to generate a sufficient PCR product out of the microdissected 20 μm slides in every tenth sample. Surprisingly, the LC, based on a clamping-PCR, was not more sensitive than sequencing (PS, Sanger). A clear advantage in sensitivity could not be found for any of the applied methods.