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Article

Detection of EGFR Mutations in Plasma cfDNA and Paired CTCs of NSCLC Patients before and after Osimertinib Therapy Using Crystal Digital PCR

1
Analysis of Circulating Tumor Cells Lab, Lab of Analytical Chemistry, Department of Chemistry, National and Kapodistrian University of Athens, 15771 Athens, Greece
2
Department of Medical Oncology, General University Hospital of Larissa, 41110 Larissa, Greece
3
Hellenic Oncology Research Group (HORG), 11471 Athens, Greece
*
Author to whom correspondence should be addressed.
Academic Editor: Roberta Alfieri
Cancers 2021, 13(11), 2736; https://doi.org/10.3390/cancers13112736
Received: 5 April 2021 / Revised: 13 May 2021 / Accepted: 27 May 2021 / Published: 31 May 2021
Liquid biopsy is a useful tool during longitudinal monitoring of NSCLC patients and requires highly sensitive and reliable technologies for accurate detection of genomic alterations. We evaluated and used crystal digital PCR to detect and quantify EGFR mutations in plasma cfDNA and paired CTCs of NSCLC patients before treatment with osimertinib and at progression of disease.
Circulating tumor DNA (ctDNA) analysis has clinical utility in EGFR mutant NSCLC. Circulating tumor cells (CTCs) consist a unique source of information at the cellular level. Digital PCR (dPCR) is a valuable tool for accurate and valid analysis of gene mutations in liquid biopsy analysis. In the present study we detected EGFR mutations in ctDNA and paired CTCs under osimertinib therapy at two time points using crystal dPCR and the naica® system (Stilla Technologies). We quantified mutation allele frequencies (MAF) of EGFR mutations in 91 plasma cfDNA samples of 48 EGFR mutant NSCLC patients and in 64 matched CTC-derived genomic DNA samples, and the FDA-cleared cobas® EGFR mutation test in 80 identical plasma samples. Direct comparison between crystal dPCR and the cobas EGFR assay revealed a high concordance for all EGFR mutations. Our comparison of crystal dPCR results in ctDNA with the corresponding primary tissue has shown a strong correlation. EGFR mutations analysis in paired CTC-derived gDNA revealed a high heterogeneity. Crystal dPCR offers the unique advantages of high analytical sensitivity, precision, and accuracy for detecting and quantifying multiple EGFR mutations in plasma cfDNA and CTCs of NSCLC patients. View Full-Text
Keywords: liquid biopsy; circulating tumor DNA (ctDNA); circulating tumor cells (CTC); crystal digital PCR; EGFR mutations; osimertinib liquid biopsy; circulating tumor DNA (ctDNA); circulating tumor cells (CTC); crystal digital PCR; EGFR mutations; osimertinib
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MDPI and ACS Style

Ntzifa, A.; Kotsakis, A.; Georgoulias, V.; Lianidou, E. Detection of EGFR Mutations in Plasma cfDNA and Paired CTCs of NSCLC Patients before and after Osimertinib Therapy Using Crystal Digital PCR. Cancers 2021, 13, 2736. https://doi.org/10.3390/cancers13112736

AMA Style

Ntzifa A, Kotsakis A, Georgoulias V, Lianidou E. Detection of EGFR Mutations in Plasma cfDNA and Paired CTCs of NSCLC Patients before and after Osimertinib Therapy Using Crystal Digital PCR. Cancers. 2021; 13(11):2736. https://doi.org/10.3390/cancers13112736

Chicago/Turabian Style

Ntzifa, Aliki, Athanasios Kotsakis, Vassilis Georgoulias, and Evi Lianidou. 2021. "Detection of EGFR Mutations in Plasma cfDNA and Paired CTCs of NSCLC Patients before and after Osimertinib Therapy Using Crystal Digital PCR" Cancers 13, no. 11: 2736. https://doi.org/10.3390/cancers13112736

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