Evaluation of KRAS, NRAS and BRAF mutations detection in plasma using an automated system for patients with metastatic colorectal cancer

Autoři: Claire Franczak aff001;  Andréa Witz aff001;  Karen Geoffroy aff001;  Jessica Demange aff001;  Marie Rouyer aff001;  Marie Husson aff001;  Vincent Massard aff002;  Céline Gavoille aff002;  Aurélien Lambert aff002;  Pauline Gilson aff003;  Nicolas Gambier aff004;  Julien Scala-Bertola aff004;  Jean-Louis Merlin aff003;  Alexandre Harlé aff003
Působiště autorů: Institut de Cancérologie de Lorraine, Service de Biopathologie, Vandoeuvre les Nancy, France aff001;  Institut de Cancérologie de Lorraine, Département d’oncologie médicale, Vandoeuvre les Nancy, France aff002;  Université de Lorraine, CNRS UMR 7039 CRAN, Institut de Cancérologie de Lorraine, Service de Biopathologie, Nancy, France aff003;  CHRU Nancy, Department of Clinical Pharmacology and Toxicology, Nancy, France aff004;  Université de Lorraine, UMR 7365 CNRS-UL, IMoPA, Vandœuvre-lès-Nancy, France aff005
Vyšlo v časopise: PLoS ONE 15(1)
Kategorie: Research Article
doi: 10.1371/journal.pone.0227294



Cell-free DNA detection is becoming a surrogate assay for tumor genotyping. Biological fluids often content a very low amount of cell-free tumor DNA and assays able to detect very low allele frequency mutant with a few quantities of DNA are required. We evaluated the ability of the fully-automated molecular diagnostics platform Idylla for the detection of KRAS, NRAS and BRAF hotspot mutations in plasma from patients with metastatic colorectal cancer (mCRC).

Materials and methods

First, we evaluated the limit of detection of the system using two set of laboratory made samples that mimic mCRC patient plasma, then plasma samples from patients with mCRC were assessed using Idylla system and BEAMing digital PCR technology.


Limits of detection of 0.1%, 0.4% and 0.01% for KRAS, NRAS and BRAF respectively have been reached. With our laboratory made samples, sensitivity up to 0.008% has been reached. Among 15 patients’ samples tested for KRAS mutation, 2 discrepant results were found between Idylla and BEAMing dPCR. A 100% concordance between the two assays has been found for the detection of NRAS and BRAF mutations in plasma samples.


The Idylla system does not reach as high sensitivity as assays like ddPCR but has an equivalent sensitivity to modified NGS technics with a lower cost and a lower time to results. These data allowed to consider the Idylla system in a routine laboratory workflow for KRAS, NRAS and BRAF mutations detection in plasma.

Klíčová slova:

Blood plasma – Circulating tumor DNA – DNA – DNA extraction – DNA fragmentation – Mutation – Mutation detection – Polymerase chain reaction


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