Evaluation of KRAS, NRAS and BRAF hotspot mutations detection for patients with metastatic colorectal cancer using direct DNA pipetting in a fully-automated platform and Next-Generation Sequencing for laboratory workflow optimisation.

Date publication

janvier 2019

Journal

PloS one

Auteurs

Membres identifiés du Cancéropôle Est :
Pr HARLE Alexandre , Dr LEROUX Agnès , Pr MERLIN Jean-Louis , Dr GILSON Pauline

Tous les auteurs :
Gilson P, Franczak C, Dubouis L, Husson M, Rouyer M, Demange J, Perceau M, Leroux A, Merlin JL, Harlé A

Lien Pubmed

https://www.ncbi.nlm.nih.gov/pubmed/31265477

Résumé

Assessment of KRAS, NRAS (RAS) and BRAF mutations is a standard in the management of patients with metastatic colorectal cancer (mCRC). Mutations could be assessed using next-generation sequencing (NGS) or real-time PCR-based assays. Times to results are 1 to 2 weeks for NGS and 1 to 3 days for real-time PCR-based assays. Using NGS can delay first-line treatment commencement and using PCR-based assays is limited by the number of possible analysed targets. The Idylla system is a real-time PCR cartridge-based assay, able to analyse hotspots mutations using one section of FFPE tumour tissue sample. To combine short delays and analysis of a large gene-panel, we propose here a laboratory workflow combining the Idylla system and NGS and compatible with FFPE samples with low tissue quantity. In this study we evaluated and validated the Idylla system for the analysis of RAS and BRAF mutations by pipetting directly DNA in the cartridge instead of FFPE section as recommended by the manufacturer.

Référence

PLoS ONE. 2019 ;14(7):e0219204