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A Multicenter Study to Assess EGFR Mutational Status in Plasma: Focus on an Optimized Workflow for Liquid Biopsy in a Clinical Setting

A multicenter study was performed to determine an optimal workflow for liquid biopsy in a clinical setting. In total, 549 plasma samples from 234 non-small cell lung cancer (NSCLC) patients were collected. Epidermal Growth Factor Receptor (EGFR) circulating cell-free tumor DNA (ctDNA) mutational ana...

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Autores principales: Sorber, Laure, Zwaenepoel, Karen, De Winne, Koen, Van Casteren, Kaat, Augustus, Elien, Jacobs, Julie, Zhang, Xiang Hua, Galdermans, Daniëlla, De Droogh, Els, Lefebure, Anneke, Morel, Ann-Marie, Saenen, Erika, Bustin, Frédérique, Demedts, Ingel, Himpe, Ulrike, Pieters, Thierry, Germonpré, Paul, Derijcke, Sofie, Deschepper, Koen, Van Meerbeeck, Jan P., Rolfo, Christian, Pauwels, Patrick
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6162772/
https://www.ncbi.nlm.nih.gov/pubmed/30150518
http://dx.doi.org/10.3390/cancers10090290
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author Sorber, Laure
Zwaenepoel, Karen
De Winne, Koen
Van Casteren, Kaat
Augustus, Elien
Jacobs, Julie
Zhang, Xiang Hua
Galdermans, Daniëlla
De Droogh, Els
Lefebure, Anneke
Morel, Ann-Marie
Saenen, Erika
Bustin, Frédérique
Demedts, Ingel
Himpe, Ulrike
Pieters, Thierry
Germonpré, Paul
Derijcke, Sofie
Deschepper, Koen
Van Meerbeeck, Jan P.
Rolfo, Christian
Pauwels, Patrick
author_facet Sorber, Laure
Zwaenepoel, Karen
De Winne, Koen
Van Casteren, Kaat
Augustus, Elien
Jacobs, Julie
Zhang, Xiang Hua
Galdermans, Daniëlla
De Droogh, Els
Lefebure, Anneke
Morel, Ann-Marie
Saenen, Erika
Bustin, Frédérique
Demedts, Ingel
Himpe, Ulrike
Pieters, Thierry
Germonpré, Paul
Derijcke, Sofie
Deschepper, Koen
Van Meerbeeck, Jan P.
Rolfo, Christian
Pauwels, Patrick
author_sort Sorber, Laure
collection PubMed
description A multicenter study was performed to determine an optimal workflow for liquid biopsy in a clinical setting. In total, 549 plasma samples from 234 non-small cell lung cancer (NSCLC) patients were collected. Epidermal Growth Factor Receptor (EGFR) circulating cell-free tumor DNA (ctDNA) mutational analysis was performed using digital droplet PCR (ddPCR). The influence of (pre-) analytical variables on ctDNA analysis was investigated. Sensitivity of ctDNA analysis was influenced by an interplay between increased plasma volume (p < 0.001) and short transit time (p = 0.018). Multistep, high-speed centrifugation both increased plasma generation (p < 0.001) and reduced genomic DNA (gDNA) contamination. Longer transit time increased the risk of hemolysis (p < 0.001) and low temperatures were shown to have a negative effect. Metastatic sites were found to be strongly associated with ctDNA detection (p < 0.001), as well as allele frequency (p = 0.034). Activating mutations were detected in a higher concentration and allele frequency compared to the T790M mutation (p = 0.003, and p = 0.002, respectively). Optimization of (pre-) analytical variables is key to successful ctDNA analysis. Sufficient plasma volumes without hemolysis or gDNA contamination can be achieved by using multistep, high-speed centrifugation, coupled with short transit time and temperature regulation. Metastatic site location influenced ctDNA detection. Finally, ctDNA levels might have further value in detecting resistance mechanisms.
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spelling pubmed-61627722018-10-02 A Multicenter Study to Assess EGFR Mutational Status in Plasma: Focus on an Optimized Workflow for Liquid Biopsy in a Clinical Setting Sorber, Laure Zwaenepoel, Karen De Winne, Koen Van Casteren, Kaat Augustus, Elien Jacobs, Julie Zhang, Xiang Hua Galdermans, Daniëlla De Droogh, Els Lefebure, Anneke Morel, Ann-Marie Saenen, Erika Bustin, Frédérique Demedts, Ingel Himpe, Ulrike Pieters, Thierry Germonpré, Paul Derijcke, Sofie Deschepper, Koen Van Meerbeeck, Jan P. Rolfo, Christian Pauwels, Patrick Cancers (Basel) Article A multicenter study was performed to determine an optimal workflow for liquid biopsy in a clinical setting. In total, 549 plasma samples from 234 non-small cell lung cancer (NSCLC) patients were collected. Epidermal Growth Factor Receptor (EGFR) circulating cell-free tumor DNA (ctDNA) mutational analysis was performed using digital droplet PCR (ddPCR). The influence of (pre-) analytical variables on ctDNA analysis was investigated. Sensitivity of ctDNA analysis was influenced by an interplay between increased plasma volume (p < 0.001) and short transit time (p = 0.018). Multistep, high-speed centrifugation both increased plasma generation (p < 0.001) and reduced genomic DNA (gDNA) contamination. Longer transit time increased the risk of hemolysis (p < 0.001) and low temperatures were shown to have a negative effect. Metastatic sites were found to be strongly associated with ctDNA detection (p < 0.001), as well as allele frequency (p = 0.034). Activating mutations were detected in a higher concentration and allele frequency compared to the T790M mutation (p = 0.003, and p = 0.002, respectively). Optimization of (pre-) analytical variables is key to successful ctDNA analysis. Sufficient plasma volumes without hemolysis or gDNA contamination can be achieved by using multistep, high-speed centrifugation, coupled with short transit time and temperature regulation. Metastatic site location influenced ctDNA detection. Finally, ctDNA levels might have further value in detecting resistance mechanisms. MDPI 2018-08-27 /pmc/articles/PMC6162772/ /pubmed/30150518 http://dx.doi.org/10.3390/cancers10090290 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Sorber, Laure
Zwaenepoel, Karen
De Winne, Koen
Van Casteren, Kaat
Augustus, Elien
Jacobs, Julie
Zhang, Xiang Hua
Galdermans, Daniëlla
De Droogh, Els
Lefebure, Anneke
Morel, Ann-Marie
Saenen, Erika
Bustin, Frédérique
Demedts, Ingel
Himpe, Ulrike
Pieters, Thierry
Germonpré, Paul
Derijcke, Sofie
Deschepper, Koen
Van Meerbeeck, Jan P.
Rolfo, Christian
Pauwels, Patrick
A Multicenter Study to Assess EGFR Mutational Status in Plasma: Focus on an Optimized Workflow for Liquid Biopsy in a Clinical Setting
title A Multicenter Study to Assess EGFR Mutational Status in Plasma: Focus on an Optimized Workflow for Liquid Biopsy in a Clinical Setting
title_full A Multicenter Study to Assess EGFR Mutational Status in Plasma: Focus on an Optimized Workflow for Liquid Biopsy in a Clinical Setting
title_fullStr A Multicenter Study to Assess EGFR Mutational Status in Plasma: Focus on an Optimized Workflow for Liquid Biopsy in a Clinical Setting
title_full_unstemmed A Multicenter Study to Assess EGFR Mutational Status in Plasma: Focus on an Optimized Workflow for Liquid Biopsy in a Clinical Setting
title_short A Multicenter Study to Assess EGFR Mutational Status in Plasma: Focus on an Optimized Workflow for Liquid Biopsy in a Clinical Setting
title_sort multicenter study to assess egfr mutational status in plasma: focus on an optimized workflow for liquid biopsy in a clinical setting
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6162772/
https://www.ncbi.nlm.nih.gov/pubmed/30150518
http://dx.doi.org/10.3390/cancers10090290
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