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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...
Autores principales: | , , , , , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2018
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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. |
format | Online Article Text |
id | pubmed-6162772 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
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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