Detection of EGFR Mutations From Plasma of NSCLC Patients Using an Automatic Cartridge-Based PCR System

Abstract

The introduction of liquid biopsies for the detection of EGFR mutations in non-small cell lung cancer patients (NSCLC) has revolutionized the clinical care. However, liquid biopsies are technically challenging and require specifically trained personnel. To facilitate the implementation of liquid biopsies for the detection of EGFR mutations from plasma, we have assessed a fully automated cartridge-based qPCR test that allows the automatic detection of EGFR mutations directly from plasma. We have analyzed 54 NSCLC patients and compared the results of the cartridge-base device to an FDA-approved assay. Detection of EGFR mutations was comparable but slightly lower in the cartridge-based device for L858R mutations (14/15 detected, 93%) and exon 19 deletions (18/20 detected, 90%). Unfortunately, 8/54 (15%) tests failed but increasing the proteinase K volume helped to recover 3/4 (75%) unsuccessful samples. In summary, the fully automated cartridge-based device allowed the detection of EGFR mutations directly from plasma in NSCLC patients with promising accuracy. However, protocol adjustments are necessary to reduce a high test failure rate.

Contribution of the IdyllaTM System to Improving the Therapeutic Care of Patients with NSCLC through Early Screening of EGFR Mutations

“…The off-label use of CE-IVD methods was thoroughly validated before being used in routine testing during a retrospective study not reported here. The risk to exhaust the tumor sample could, however, be lifted by the reuse of H&E, immunohistochemistry (IHC), and fluorescence in situ hybridization (FISH) diagnostic slides [ 36 ], by the use of plasma from patients with lung cancer [ 37 , 38 ] or of the DNA extracted from FFPE sections for NGS analysis [ 14 , 19 , 26 , 28 , 39 ]. Indeed, using the same DNA for Idylla TM and NGS assays could discard divergent results linked to tumor heterogeneity and to the fact that the two analyses are not carried out on strictly the same part of the tumor sample.…”

Machine Learning-Based Radiomics Signatures for EGFR and KRAS Mutations Prediction in Non-Small-Cell Lung Cancer

“…Epidermal growth factor receptor (EGFR) and Kirsten rat sarcoma viral oncogene homolog (KRAS) are the two most frequently observed mutations in patients with NSCLC [4]. Patients with EGFR mutations exhibit higher sensitivity to gefitinib and erlotinib, whereas those with KRAS mutations are prone to drug resistance [4][5][6][7][8][9]. Thus, the early detection of EGFR and KRAS mutations can facilitate the selection of treatment modalities for specific mutation types and enable efficient short-and long-term management, thus improving the prognosis and prolonging the survival duration of patients with NSCLC [4].…”

Daily Practice Assessment of KRAS Status in NSCLC Patients: A New Challenge for the Thoracic Pathologist Is Right around the Corner

“…However, NGS with a LB is currently not often available in most laboratories, in Europe at least, and is set up for routine testing in only a few comprehensive cancer centers [ 191 , 215 , 218 ]. Therefore, many laboratories use targeted sequencing methods, such as RT-PCR or ddPCR [ 219 , 220 , 221 ]. Interestingly, a recent study demonstrated that the amount of cf-DNA was higher when some genomic alterations such as KRAS were identified in tumors, in contrast to a lower amount of cf-DNA associated with some other mutations detected on EGFR or TP53 [ 222 ].…”