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Open AccessArticle

Detection of EGFR Mutations in Plasma Cell-Free Tumor DNA of TKI-Treated Advanced-NSCLC Patients by Three Methodologies: Scorpion-ARMS, PNAClamp, and Digital PCR

1
Medical Oncology Division, S. Maria della Misericordia Hospital, 06132 Perugia, Italy
2
Biosciences Laboratory, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, 47014 Meldola, Italy
3
Department of Medical Oncology, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, 47014 Meldola, Italy
*
Author to whom correspondence should be addressed.
These authors contributed equally to the work.
Diagnostics 2020, 10(12), 1062; https://doi.org/10.3390/diagnostics10121062
Received: 4 November 2020 / Revised: 4 December 2020 / Accepted: 4 December 2020 / Published: 7 December 2020
Analysis of circulating cell-free tumor DNA (cftDNA) has emerged as a specific and sensitive blood-based approach to detect epidermal growth factor receptor (EGFR) mutations in non-small cell lung cancer (NSCLC) patients. Still, there is some debate on what should be the preferential clinical method for plasma-derived cftDNA analysis. We tested 31 NSCLC patients treated with anti-EGFR tyrosine kinase inhibitors (TKIs), at baseline and serially during therapy, by comparing three methodologies in detecting EGFR mutations (L858R, exon 19 deletion, and T790M) from plasma: scorpions-amplification refractory mutation system (ARMS) methodology by using EGFR Plasma RGQ PCR Kit-QIAGEN, peptide nucleic acid (PNA) clamp and PANA RealTyper integration by using PNAClamp EGFR-PANAGENE, and digital real time PCR by using QuantStudio 3D Digital PCR System-Thermo Fisher Scientific. Specificity was 100% for all three mutations, independently from the platform used. The sensitivity for L858R (42.86%) and T790M (100%) did not change based on the method, while the sensitivity for Del 19 differed markedly (Scorpion-ARMS 45%, PNAClamp 75%, and Digital PCR 85%). The detection rate was also higher (94.23%) as measured by Digital PCR, and when we monitored the evolution of EGFR mutations over time, it evidenced the extreme inter-patient heterogeneity in terms of levels of circulating mutated copies. In our study, Digital PCR showed the best correlation with tissue biopsy and the highest sensitivity to attain the potential clinical utility of monitoring plasma levels of EGFR mutations. View Full-Text
Keywords: EGFR; TKIs; cftDNA; liquid biopsy; NSCLC EGFR; TKIs; cftDNA; liquid biopsy; NSCLC
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MDPI and ACS Style

Siggillino, A.; Ulivi, P.; Pasini, L.; Reda, M.S.; Chiadini, E.; Tofanetti, F.R.; Baglivo, S.; Metro, G.; Crinó, L.; Delmonte, A.; Minotti, V.; Roila, F.; Ludovini, V. Detection of EGFR Mutations in Plasma Cell-Free Tumor DNA of TKI-Treated Advanced-NSCLC Patients by Three Methodologies: Scorpion-ARMS, PNAClamp, and Digital PCR. Diagnostics 2020, 10, 1062. https://doi.org/10.3390/diagnostics10121062

AMA Style

Siggillino A, Ulivi P, Pasini L, Reda MS, Chiadini E, Tofanetti FR, Baglivo S, Metro G, Crinó L, Delmonte A, Minotti V, Roila F, Ludovini V. Detection of EGFR Mutations in Plasma Cell-Free Tumor DNA of TKI-Treated Advanced-NSCLC Patients by Three Methodologies: Scorpion-ARMS, PNAClamp, and Digital PCR. Diagnostics. 2020; 10(12):1062. https://doi.org/10.3390/diagnostics10121062

Chicago/Turabian Style

Siggillino, Annamaria; Ulivi, Paola; Pasini, Luigi; Reda, Maria S.; Chiadini, Elisa; Tofanetti, Francesca R.; Baglivo, Sara; Metro, Giulio; Crinó, Lucio; Delmonte, Angelo; Minotti, Vincenzo; Roila, Fausto; Ludovini, Vienna. 2020. "Detection of EGFR Mutations in Plasma Cell-Free Tumor DNA of TKI-Treated Advanced-NSCLC Patients by Three Methodologies: Scorpion-ARMS, PNAClamp, and Digital PCR" Diagnostics 10, no. 12: 1062. https://doi.org/10.3390/diagnostics10121062

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