Comparison of PNA Clamping-assisted Fluorescence Melting Curve Analysis and PNA Clamping in Detecting EGFR Mutations in Matched Tumor Tissue, Cell Block, Pleural Effusion and Blood of Lung Cancer Patients With Malignant Pleural Effusion

Abstract

Background/Aim: This study compared the efficacy of PANAMutyper™, a novel technology that integrates PNAClamp™ and PANA S-Melting™, and PNAClamp™ alone for the detection of EGFR mutations in lung cancer patients. Materials and Methods: PANAMutyper™ and PNAClamp™ were used to assess the EGFR mutation status in tissue, cell block, pleural effusion, and blood samples of 90 lung cancer patients with malignant pleural effusion. Results: PANAMutyper™ detected more EGFR mutations than PNAClamp™, especially in body fluids (pleural effusion and serum). Patients with additional EGFR mutations detected using PANAMutyper™ had a favorable response to EGFRtyrosine kinase inhibitor (TKI) treatment. Conclusion: The diagnostic performance of PANAMutyper™ was superior to that of PNAClamp™ for the detection of EGFR mutations. It was also better at identifying lung cancer patients with malignant pleural effusion who were likely to benefit from EGFR-TKI treatment.The use of molecular agents targeting the epidermal growth factor receptor (EGFR) is important in the treatment of advanced non-small cell lung cancer (NSCLC) (1-3).Multiple prospective clinical trials have demonstrated that patients with advanced NSCLC harboring activating mutations in the EGFR gene show improved objective response rates and progression-free survival (PFS) when treated with EGFR tyrosine kinase inhibitors (EGFR-TKIs) (1,2,(4)(5)(6)(7)(8). Therefore, molecular testing for EGFR mutations has become essential for predicting whether a patient will benefit from EGFR-TKI targeted therapy (9).Previously, we compared peptide nucleic acid (PNA) clamping with direct sequencing for the detection of EGFR and K-RAS mutations. Our results showed that PNA clamping has a better diagnostic performance and higher clinical significance (10, 11).

Evaluation of molecular methods for plasma detection of EGFR mutations in non‐small cell lung cancer

“…Overall sensitivity of the Cobas (76.7%) was higher than that of the PANAMutyper (60.5%) in this study. In particular, the most common EGFR mutation, exon 19 deletions, was better detected by the Cobas than by the other methods, similar to a prior study 21 . The detection rates of the L858R and T790M did not differ significantly among the three plasma‐based tests.…”

“…The Cobas was used to detect EGFR mutations in cytology-negative samples and increase the diagnostic yield. 32 Jeon et al 21 reported good diagnostic performance of the PANAMutyper This study has some limitations. First, the timing of each test using plasma was different; there may be an effect of storage duration.…”

“…In particular, the most common EGFR mutation, exon 19 deletions, was better detected by the Cobas than by the other methods, similar to a prior study. 21 The detection rates of the L858R and T790M did not differ significantly among the three plasma-based tests. Based on the tissue results, the Cobas and PANAMutyper displayed overall specificities of 92%∼96%, similar to the 90%∼100% specificities reported previously.…”

“…The Cobas was used to detect EGFR mutations in cytology‐negative samples and increase the diagnostic yield 32 . Jeon et al 21 . reported good diagnostic performance of the PANAMutyper with pleural fluids compared to tissues or cell blocks.…”

Programmed death-ligand 1 expression level as a predictor of EGFR tyrosine kinase inhibitor efficacy in lung adenocarcinoma

“…EGFR gene mutations were analyzed by a peptide nucleic acid (PNA) clamping-based sensitive method (PNA Clamp™ or PANAMutyper™ EGFR mutation detection kit) in formalin-fixed paraffin-embedded tissues (16)(17)(18). EGFR-activating mutations were defined as mutations associated with EGFR-TKI sensitivity, including exon 19 deletion, exon 21 L858R, and L861Q.…”