High Resolution Melting Analysis for Epidermal Growth Factor Receptor Mutations in Formalin-fixed Paraffin-embedded Tissue and Plasma Free DNA from Non-small Cell Lung Cancer Patients

Background:The aim of the research was to explore a cost effective, fast, easy to perform, and sensitive methodfor epidermal growth factor receptor (EGFR) mutation testing. Methods: High resolution melting analysis (HRM)was introduced to evaluate the efficacy of the analysis for dectecting EGFR mutations in exons 18 to 21 usingformalin-fixed paraffin-embedded (FFPE) tissues and plasma free DNA from 120 patients. Results: The totalEGFR mutation rate was 37.5% (45/120) detected by direct sequencing. There were 48 mutations in 120 FFPEtissues assessed by HRM. For plasma free DNA, the EGFR mutation rate was 25.8% (31/120). The sensitivityof HRM assays in FFPE samples was 100% by HRM. There was a low false-positive mutation rate but a highfalse-negative rate in plasma free DNA detected by HRM. Conclusions: Our results show that HRM analysis hasthe advantage of small tumor sample need. HRM applied with plasma free DNA showed a high false-negativerate but a low false-positive rate. Further research into appropriate methods and analysis needs to be performedbefore HRM for plasma free DNA could be accepted as an option in diagnostic or screening settings.