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
Résumé
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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