Assessment of Common Somatic Mutations of EGFR,KRAS,BRAF,NRAS and PIK3CA in Pulmonary Adenocarcinoma using iPLEXHS,a New Highly Sensitive Assay for MassARRAY.

Increased early detection and personalized therapy for lung cancer have coincided with greater use of minimally invasive sampling techniques such as endobronchial ultrasound-guided biopsy (EBUS),endoscopic ultrasound-guided biopsy (EUS),and navigational biopsy,as well as thin needle core biopsies. As many lung cancer patients have late stage disease and other comorbidities that make open surgical procedures hazardous,the least invasive biopsy technique with the highest potential specimen yield is now the preferred first diagnostic study. However,use of these less invasive procedures generates significant analytical challenges for the laboratory,such as a requirement for robust detection of low level somatic mutations,particularly when the starting sample is very small or demonstrates few intact tumor cells. In this study,we assessed 179 clinical cases of non-small cell lung carcinoma (NSCLC) that had been previously tested for EGFR,KRAS,NRAS,and BRAF mutations using a novel multiplexed analytic approach that reduces wild-type signal and allows for detection of low mutation load approaching 1%,iPLEX® HS panel for the MassARRAY® System (Agena Bioscience,San Diego,CA). This highly sensitive system identified approximately 10% more KRAS,EGFR and BRAF mutations than were detected by the original test platform,which had a sensitivity range of 5-10% variant allele frequency (VAF).