TY - JOUR T1 - Validation of PNA-LNA PCR clamp assay for detection of EGFR exon 19 and 21 mutations in various types of clinical non-small cell lung cancer specimens JF - European Respiratory Journal JO - Eur Respir J VL - 42 IS - Suppl 57 SP - P544 AU - Michal Skronski AU - Paulina Jagus AU - Renata Langfort AU - Adam Szpechcinski AU - Krystyna Maszkowska-Kopij AU - Barbara Roszkowska-Sliz AU - Jacek Grudny AU - Jolanta Zaleska AU - Tadeusz Orlowski AU - Kazimierz Roszkowski-Sliz AU - Joanna Chorostowska-Wynimko Y1 - 2013/09/01 UR - http://erj.ersjournals.com/content/42/Suppl_57/P544.abstract N2 - The aim of the study was to assess diagnostic reliability of PNA-LNA PCR clamp assay in EGFR mutations detection in different NSCLC samples.Evaluation was performed: (i) in reference NSCLC tissue FFPE samples (n=10), (ii) in comparison to direct sequencing in resected NSCLC tissue (n=199) and biopsy material specimens (n=179) characterized by different tumor cells content (TCC) and fixation [Table 1].View this table:NSCLC samples characteristic.(i) PNA-LNA PCR clamp correctly detected all exon 19 deletions and L858R mutations in the reference FFPE materials, including those with meager TCC (5% and 10%). (ii) PNA-LNA PCR clamp method and direct sequencing presented high conformity (overall percent agreement, OPA=99%; Cohen’s Kappa score of 0.94 (95% CI=0.9, 0.99) in n=100 samples with >50% TCC analyzed. (iii) In total of 378 samples analyzed with PNA-LNA PCR clamp method, EGFR mutations were detected in 36 (9.5%). (iv) Only in 24 out of 36 (67%) mutation positive samples reevaluation with direct sequencing proved positive [Table 2]. PNA-LNA PCR clamp presented higher sensitivity in samples with TCC <50% (p=0.004).View this table:Comparison of PNA-LNA PCR clamp vs direct sequencing EGFR mutation detection sensitivity in materials with different TCC.PNA-LNA PCR clamp method proved its diagnostic utility and high sensitivity of EGFR exon 19 and 21 mutations detection, especially in samples with TCC lower than 50%. ER -