Assessment of KRAS status is mandatory in patients with metastatic colorectal cancer (mCRC) before applying targeted therapy. We describe here a blinded prospective study to compare KRAS and BRAF mutation status data obtained from the analysis of tumor tissue by routine gold-standard methods and of plasma DNA using a quantitative PCR–based method specifically designed to analyze circulating cell-free DNA (cfDNA). The mutation status was determined by both methods from 106 patient samples. cfDNA analysis showed 100% specificity and sensitivity for the BRAF V600E mutation. For the seven tested KRAS point mutations, the method exhibited 98% specificity and 92% sensitivity with a concordance value of 96%. Mutation load, expressed as the proportion of mutant alleles in cfDNA, was highly variable (0.5–64.1%, median 10.5%) among mutated samples. CfDNA was detected in 100% of patients with mCRC. This study shows that liquid biopsy through cfDNA analysis could advantageously replace tumor-section analysis and expand the scope of personalized medicine for patients with cancer.
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We thank J. Robert, J. Dubois, A. Lievre, C. Theillet, S. Du Manoir, C. Sardet, D. Tousch and R. Gimbaud for helpful discussions. We would like to thank L. Kamraoui, A. Laybats, J.-Y. Cance and M. Cavalier for excellent technical assistance. F. Mouliere is supported by a grant from CNRS and the Region of Languedoc-Roussillon (CNRS044406). The study was funded by grants from the Fondation ARC pour la Recherche sur le Cancer and the Association GEFLUC (DCMLP10-184, France). We thank Bio-Rad for the qPCR thermocycler free loan. A.R.T. is supported by INSERM.
The authors declare no competing financial interests.
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Thierry, A., Mouliere, F., El Messaoudi, S. et al. Clinical validation of the detection of KRAS and BRAF mutations from circulating tumor DNA. Nat Med 20, 430–435 (2014). https://doi.org/10.1038/nm.3511
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