Abstract
The aim of our study was to evaluate the effectiveness of EGFR gene mutation detection in diverse materials from 707 NSCLC patients using the two ultra-sensitive allele-specific real-time PCR assays in the routine diagnostic procedure.
629 samples were analyzed by PNA-LNA PCR clamp, 164 samples by CE-IVD certified allele-specific PCR assay and 86 samples by both methods. PNA-LNA PCR clamp products were analyzed by Sanger sequencing to confirm rare mutations. NSCLC specimens were characterized by tumor cells content and type of fixation
NSCLC samples
.
62/707 (9%) specimens were positive for EGFR mutations: 32 detected in exon 19, including 4 rare deletions and p.R748K substitution (c.2243G>A); 28 p.L858R (c.2573T>G) mutations, 1 rare variant of p.L858R (c.2572_2573delinsAG) and 1 double mutation p.L858M+p.L861Q (c.2572C>A; c.2582T>A) detected in exon 21. There were 71% women in the EGFR+ group. 98% of EGFR+ were adenocarcinoma samples, thus mutation frequency among adenocarcinoma patients was 9.8% (61/617 patients). Ultrasensitive PNA-LNA PCR clamp assay and allele-specific PCR CE-IVD test presented high results conformity with 95.3% overall percent agreement (95%CI=90.9-99.8) and Cohen's Kappa score of 0.9 (95%CI=0.88-0.92).
Both real-time PCR assays provided reliable and robust detection of most common EGFR activating mutations in various clinical NSCLC specimens.
- © 2014 ERS
