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. 2019 Aug 12;9:11610. doi: 10.1038/s41598-019-47489-7

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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(A) Sample Workflow. This flowchart explains the workflow used in this study starting from whole blood samples collected in this study to the analysis performed. (B) Combined copy number and mutational analysis of cfDNA. Combined mutational and copy number plot for the 85 genes that were positive for at least 1 mutation across all 55 cfDNA samples analysed. Boxes coloured orange represent mutation and a copy number gain (CNG), yellow mutation and copy number loss (CNL), green mutation, red CNG, and blue for CNL. (C) Kaplan-Meier analysis of the overall survival according to KRAS mutation alone (7/55), KRAS mutation and copy number gain (7/55), along with KRAS wild-type (41/55) in 55 patients revealed best prognosis for patients with KRAS wild-type, with a median survival of 10.6 months, followed by patients with KRAS mutation alone with a median survival of 5.6 months. Patients with both the KRAS mutation and copy number gain had the worst prognosis with a median survival of 2.5 months (overall Log-rank test p-value = < 0.0001). KRAS WT vs KRAS MUT only Log-rank p-value = 0.0610, KRAS WT vs KRAS CN gain + MUT Log-rank p = value = 0.0012 and KRAS MUT only vs KRAS CN gain + MUT Log-rank p-value = < 0.0001. (D) Hazard ratios for the factors that were used in a multivariable analysis.