Detection of KRAS mutation in colorectal cancer patients' cfDNA with droplet digital PCR.
- Author:
Yuwen LUO
1
;
Yao LI
1
Author Information
- Publication Type:Journal Article
- Keywords: KRAS; cell free DNA; droplet digital PCR colorectal cancer; liquid biopsy
- From: Chinese Journal of Biotechnology 2018;34(3):407-420
- CountryChina
- Language:Chinese
- Abstract: This study aims to develop a new method for the detection of KRAS mutations related to colorectal cancer in cfDNA, and to evaluate the sensitivity and accuracy of the detection. We designed a method of cfDNA based KRAS detection by droplets digital PCR (ddPCR). The theoretical performance of the method is evaluated by reference standard and compared to the ARMS PCR method. Two methods, ddPCR and qPCR, were successfully established to detect KRAS wild type and 7 mutants. Both methods were validated using plasmid standards and actual samples. The results were evaluated by false positive rate, linearity, and limit of detection. Finally, 52 plasma cfDNA samples from patients and 20 samples from healthy people were tested, the clinical sensitivity is 97.64%, clinical specificity is 81.43%. ddPCR method shows higher performance than qPCR. The LOD of ddPCR method reached single digits of cfDNA copies, it can detect as low as 0.01% to 0.04% mutation abundance.
