Assessment of High-Sensitive Methods for the Detection of EGFR Mutations in Circulating Free Tumor DNA from NSCLC Patients

Abstract

Aim: We assessed the ability of the Therascreen^® kit (plasma-Therascreen) and of a peptide nucleic acids (PNA)-clamp approach to detect EGFR mutations in plasma-derived circulating-free tumor DNA (cftDNA) from non-small-cell lung cancer patients. Materials & methods: cftDNA from 96 patients was analyzed for exon 19 deletions and the p.L858R mutation, using both plasma-Therascreen and PNA-clamp-based assays. Results: None of the 70 EGFR wild-type patients showed EGFR mutations in cftDNA with both techniques (specificity: 100%). In 17/26 EGFR-mutant patients, plasma-Therascreen analysis confirmed the mutation identified in the primary tumor (analytical sensitivity: 65.4%). Similar results were obtained with the PNA-clamp method. Conclusion: Both approaches were specific and sensitive for EGFR mutational analysis of cftDNA in non-small-cell lung cancer patients.

Keywords::

• cftDNA
• EGFR mutations
• liquid biopsy
• non-small-cell lung carcinoma
• PNA-clamp
• Therascreen^®

Financial disclosure & competing interests disclosure

N Normanno received honoraria from AstraZeneca, Qiagen, Roche Diagnostics as a member of speakers’ bureau. N Normanno was supported by grants from Associazione Italiana per la Ricerca sul Cancro (AIRC). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.

Ethical conduct of research

The authors state that they have obtained appropriate institutional review board approval or have followed the principles outlined in the Declaration of Helsinki for all human or animal experimental investigations. In addition, for investigations involving human subjects, informed consent has been obtained from the participants involved.

Additional information

Funding

N Normanno received honoraria from AstraZeneca, Qiagen, Roche Diagnostics as a member of speakers’ bureau. N Normanno was supported by grants from Associazione Italiana per la Ricerca sul Cancro (AIRC). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed. No writing assistance was utilized in the production of this manuscript.