Carrier molecules and extraction of circulating tumor DNA for next generation sequencing in colorectal cancer
Jazyk angličtina Země Česko Médium print
Typ dokumentu časopisecké články
PubMed
27526306
DOI
10.14712/18059694.2016.54
PII: 18059694.2016.54
Knihovny.cz E-zdroje
- Klíčová slova
- Carrier, Circulating tumor DNA, Extraction, Next generation sequencing, Real-time PCR,
- MeSH
- DNA nádorová krev genetika izolace a purifikace MeSH
- kolorektální nádory krev diagnóza genetika patologie MeSH
- kvantitativní polymerázová řetězová reakce MeSH
- lidé středního věku MeSH
- lidé MeSH
- metastázy nádorů MeSH
- mutační analýza DNA MeSH
- prognóza MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- transportní proteiny krev MeSH
- vysoce účinné nukleotidové sekvenování * MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- DNA nádorová MeSH
- transportní proteiny MeSH
The aims of the study were: i) to compare circulating tumor DNA (ctDNA) yields obtained by different manual extraction procedures, ii) to evaluate the addition of various carrier molecules into the plasma to improve ctDNA extraction recovery, and iii) to use next generation sequencing (NGS) technology to analyze KRAS, BRAF, and NRAS somatic mutations in ctDNA from patients with metastatic colorectal cancer. Venous blood was obtained from patients who suffered from metastatic colorectal carcinoma. For plasma ctDNA extraction, the following carriers were tested: carrier RNA, polyadenylic acid, glycogen, linear acrylamide, yeast tRNA, salmon sperm DNA, and herring sperm DNA. Each extract was characterized by quantitative real-time PCR and next generation sequencing. The addition of polyadenylic acid had a significant positive effect on the amount of ctDNA eluted. The sequencing data revealed five cases of ctDNA mutated in KRAS and one patient with a BRAF mutation. An agreement of 86% was found between tumor tissues and ctDNA. Testing somatic mutations in ctDNA seems to be a promising tool to monitor dynamically changing genotypes of tumor cells circulating in the body. The optimized process of ctDNA extraction should help to obtain more reliable sequencing data in patients with metastatic colorectal cancer.
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