Journal
MOLECULAR ONCOLOGY
Volume 8, Issue 5, Pages 927-941Publisher
WILEY
DOI: 10.1016/j.molonc.2014.02.005
Keywords
Circulating cell-free DNA; Multi-marker analysis; qPCR; Colorectal cancer
Categories
Funding
- Fondation ARC pour la Recherche sur le Cancer
- GEFLUC (France) [DCMLP10-184]
- Institut National de la Sante et de la Recherche Medicale (INSERM) (France)
Ask authors/readers for more resources
Development of a Q-PCR-based assay for the high-performance analysis of circulating cell-free DNA (ccfDNA) requires good knowledge of its structure and size. In this work, we present the first visual determination of ccfDNA by Atomic Force Microscopy (AFM) on plasma samples from colorectal cancer (CRC) patients and healthy donors. In addition to the examination of fragment size distribution profile as performed by Q-PCR, this analysis confirms that ccfDNA is highly fragmented and that more than 80% of ccfDNA fragments in CRC plasma are below 145 bp. We adapted an Allele-Specific Blocker (ASB) Q-PCR to small ccfDNA fragments to determine simultaneously the total ccfDNA concentration, the presence of point mutation, the proportion of mutated allele, and a ccfDNA integrity index. The data validated analytically these four parameters in 124 CRC clinical samples and 71 healthy individuals. The multi-marker method, termed Intplex, enables sensitive and specific non-invasive analysis of tumor ccfDNA, which has great potential in terms of cost, quality control, and easy implementation in every clinical center laboratory. (C) 2014 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available