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Comparison of the quantification of KRAS mutations by digital PCR and E-ice-COLD-PCR in circulating-cell-free DNA from metastatic colorectal cancer patients

Abstract : Circulating cell-free DNA (ccfDNA) bears great promise as biomarker for personalized medicine, but ccfDNA is present only at low levels in the plasma or serum of cancer patients. E-ice-COLD-PCR is a recently developed enrichment method to detect and identify mutations present at low-abundance in clinical samples. However, recent studies have shown the importance to accurately quantify low-abundance mutations as clinically important decisions will depend on certainmutation thresholds. The possibility for an enrichmentmethod to accurately quantify the mutation levels remains a point of concern and might limit its clinical applicability. In the present study, we compared the quantification of KRAS mutations in ccfDNA from metastatic colorectal cancer patients by E-ice-COLD-PCR with two digital PCR approaches. For the quantification of mutations by Eice- COLD-PCR, cell lines with known mutations diluted into WT genomic DNA were used for calibration. E-ice- COLD-PCR and the two digital PCR approaches showed the same range of the mutation level and were concordant for mutation levels below the clinical relevant threshold. E-ice-COLD-PCR can accurately detect and quantify low-abundantmutations in ccfDNA and has a shorter time to results making it compatible with the requirements of analyses in a clinical setting without the loss of quantitative accuracy.
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https://hal-normandie-univ.archives-ouvertes.fr/hal-02353197
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Submitted on : Tuesday, May 31, 2022 - 8:40:28 PM
Last modification on : Friday, June 10, 2022 - 3:31:19 PM

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David Sefrioui, Florence Mauger, Laurence Leclere, Ludivine Beaussire, Frédéric Di Fiore, et al.. Comparison of the quantification of KRAS mutations by digital PCR and E-ice-COLD-PCR in circulating-cell-free DNA from metastatic colorectal cancer patients. Clinica Chimica Acta, Elsevier, 2017, 465, pp.1-4. ⟨10.1016/j.cca.2016.12.004⟩. ⟨hal-02353197⟩

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