TY  - JOUR
AU  - Prosenc-Zmrzljak, Ursula
AU  - Kosir, Rok
AU  - Krivokapić, Zoran
AU  - Radojković, Dragica
AU  - Nikolić, Aleksandra
PY  - 2021
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1494
AB  - Liquid biopsy and cell-free DNA (cfDNA) show great promise in cancer diagnostics. In this study, we designed a custom droplet digital PCR (ddPCR) assay for the quantification and quality control of cfDNA isolated from serum. The assay was validated on a group of locally advanced colorectal cancer (CRC) patients and two control groups-patients with hemorrhoids and healthy individuals. The assay shows a high correlation with Qubit measurement (r = 0.976) but offers a higher dynamic range. Mean concentrations of cfDNA were 12.36 ng/mu L, 5.17 ng/mu L, and 0.29 ng/mu L for CRC, hemorrhoid patients, and healthy controls, respectively. The quality of cfDNA was assessed with the measurement of B-cell DNA contamination. On a subset of CRC patients, we compared the mutation status on KRAS (G12A, G12D, G12V, G13D) and BRAF (V600E) genes in the primary tumor and cfDNA isolated from the serum. A total of 70.6% of primary tumor samples were mutated, and the mean fractional abundance of mutations was 9.50%. The matching serum samples were mutated in 38% cases with an average fractional abundance of 0.23%. We conclude that any decisions based solely on the amount of cfDNA present in patient serum must be interpreted carefully and in the context of co-morbidities. This study explores the potential of ddPCR somatic mutations detection from liquid biopsy as a supplement to tissue biopsy in targeted personalized CRC patient management.
PB  - MDPI, Basel
T2  - Genes
T1  - Detection of Somatic Mutations with ddPCR from Liquid Biopsy of Colorectal Cancer Patients
IS  - 2
VL  - 12
DO  - 10.3390/genes12020289
ER  - 

@article{
author = "Prosenc-Zmrzljak, Ursula and Kosir, Rok and Krivokapić, Zoran and Radojković, Dragica and Nikolić, Aleksandra",
year = "2021",
abstract = "Liquid biopsy and cell-free DNA (cfDNA) show great promise in cancer diagnostics. In this study, we designed a custom droplet digital PCR (ddPCR) assay for the quantification and quality control of cfDNA isolated from serum. The assay was validated on a group of locally advanced colorectal cancer (CRC) patients and two control groups-patients with hemorrhoids and healthy individuals. The assay shows a high correlation with Qubit measurement (r = 0.976) but offers a higher dynamic range. Mean concentrations of cfDNA were 12.36 ng/mu L, 5.17 ng/mu L, and 0.29 ng/mu L for CRC, hemorrhoid patients, and healthy controls, respectively. The quality of cfDNA was assessed with the measurement of B-cell DNA contamination. On a subset of CRC patients, we compared the mutation status on KRAS (G12A, G12D, G12V, G13D) and BRAF (V600E) genes in the primary tumor and cfDNA isolated from the serum. A total of 70.6% of primary tumor samples were mutated, and the mean fractional abundance of mutations was 9.50%. The matching serum samples were mutated in 38% cases with an average fractional abundance of 0.23%. We conclude that any decisions based solely on the amount of cfDNA present in patient serum must be interpreted carefully and in the context of co-morbidities. This study explores the potential of ddPCR somatic mutations detection from liquid biopsy as a supplement to tissue biopsy in targeted personalized CRC patient management.",
publisher = "MDPI, Basel",
journal = "Genes",
title = "Detection of Somatic Mutations with ddPCR from Liquid Biopsy of Colorectal Cancer Patients",
number = "2",
volume = "12",
doi = "10.3390/genes12020289"
}

Prosenc-Zmrzljak, U., Kosir, R., Krivokapić, Z., Radojković, D.,& Nikolić, A.. (2021). Detection of Somatic Mutations with ddPCR from Liquid Biopsy of Colorectal Cancer Patients. in Genes
MDPI, Basel., 12(2).
https://doi.org/10.3390/genes12020289

Prosenc-Zmrzljak U, Kosir R, Krivokapić Z, Radojković D, Nikolić A. Detection of Somatic Mutations with ddPCR from Liquid Biopsy of Colorectal Cancer Patients. in Genes. 2021;12(2).
doi:10.3390/genes12020289 .

Prosenc-Zmrzljak, Ursula, Kosir, Rok, Krivokapić, Zoran, Radojković, Dragica, Nikolić, Aleksandra, "Detection of Somatic Mutations with ddPCR from Liquid Biopsy of Colorectal Cancer Patients" in Genes, 12, no. 2 (2021),
https://doi.org/10.3390/genes12020289 . .