Testing for 29 EGFR TKI Sensitivity and Resistance Mutations in Lung Cancer Using EGFR RGQ PCR Kit
Milena Cankovic, Lisa Whiteley, Dhananjay A Chitale. Henry Ford Hospital, Detroit, MI
Background: Many studies have demonstrated the association between somatic mutations in the epidermal growth factor receptor (EGFR) gene in non-small cell lung cancer (NSCLC) and sensitivity or resistance to the EGFR tyrosine kinase inhibitors (TKIs). In this study we evaluated performance of a commercial kit (EGFR RGQ PCR, Qiagen) for detection of EGFR mutations in a variety of sample types likely to be submitted to a molecular oncology lab.
Design: Qiagen's EGFR RGQ kit detects 29 somatic mutations in the EGFR oncogene using real time PCR and Rotor Gene Q (RGQ) instrument. The kit uses Scorpions and ARMS® technologies to detect these mutations in the background of wild type genomic DNA. Sixty two confirmed NSCLC cases were tested in parallel using EGFR RGQ PCR and laboratory developed assays (LDA) EGFR exon 19 deletion/exon 21 L858R point mutation. Sample types were as follows: 42 lung biopsies/resections, 3 lymph node biopsies, 3 brain biopsies/resections, 3 liver biopsies, 1 gastric biopsy, 1 skin biopsy, 4 pleural cytology fluid cell blocks, 1 pleural cytology slide smear, 2 pericardial fluid cytology cell blocks, 1 scapula resection, 1 paraspinal mass. Manual microdissection was performed when tumor content was less than 50% of total cells (20/62 cases).